Benzothiophene compounds, intermediates, compositions, and method for inhibiting restenosis

ABSTRACT

The present invention provides pharmaceutically active compounds of formula I ##STR1## wherein R 1  is --H, --OH, --O(C 1  -C 4  alkyl), --OCOC 6  H 5 , --OCO(C 1  -C 6  alkyl), or --OSO 2  (C 2  -C 6  alkyl); 
     R 2  is --H, --OH, --O(C 1  -C 4  alkyl), --OCOC 6  H 5 , --OCO(C 1  -C 6  alkyl), --OSO 2  (C 2  -C 6  alkyl), or halo, providing when Z is --S--, R 2  is not halo; 
     R 3  is 1-piperidinyl, 1-pyrrolidinyl, methyl-1-pyrrolidinyl, dimethyl-1-pyrrolidinyl, 4-morpholino, dimethylamino, diethylamino, diisopropylamino, or 1-hexamethyleneimino; 
     n is 2 or 3; and 
     z is --O-- or --S--; 
     or a pharmaceutically acceptable salt thereof, for inhibiting restenosis.

This application is a division of pending prior application Ser. No.08/396,401, filed on Feb. 28, 1995.

FIELD OF THE INVENTION

This invention relates to the fields of pharmaceutical and organicchemistry and provides novel benzothiophene compounds which are usefulfor the treatment of the various medical indications associated withpost-menopausal syndrome, and uterine fibroid disease, endometriosis,and aortal smooth muscle cell proliferation. The present inventionfurther relates to intermediate compounds useful for preparing thepharmaceutically active compounds of the present invention, andpharmaceutical compositions.

BACKGROUND OF THE INVENTION

"Post-menopausal syndrome" is a term used to describe variouspathological conditions which frequently affect women who have enteredinto or completed the physiological metamorphosis known as menopause.Although numerous pathologies are contemplated by the use of this term,three major effects of post-menopausal syndrome are the source of thegreatest long-term medical concern: osteoporosis, cardiovascular effectssuch as hyperlipidemia, and estrogen-dependent cancer, particularlybreast and uterine cancer.

Osteoporosis describes a group of diseases which arise from diverseetiologies, but which are characterized by the net loss of bone mass perunit volume. The consequence of this loss of bone mass and resultingbone fracture is the failure of the skeleton to provide adequatestructural support for the body. One of the most common types ofosteoporosis is that associated with menopause. Most women lose fromabout 20% to about 60% of the bone mass in the trabecular compartment ofthe bone within 3 to 6 years after the cessation of menses. This rapidloss is generally associated with an increase of bone resorption andformation. However, the resorptive cycle is more dominant and the resultis a net loss of bone mass. Osteoporosis is a common and serious diseaseamong post-menopausal women.

There are an estimated 25 million women in the United States, alone, whoare afflicted with this disease. The results of osteoporosis arepersonally harmful and also account for a large economic loss due itschronicity and the need for extensive and long term support(hospitalization and nursing home care) from the disease sequelae. Thisis especially true in more elderly patients. Additionally, althoughosteoporosis is not generally thought of as a life threateningcondition, a 20% to 30% mortality rate is related with hip fractures inelderly women. A large percentage of this mortality rate can be directlyassociated with post-menopausal osteoporosis.

The most vulnerable tissue in the bone to the effects of post-menopausalosteoporosis is the trabecular bone. This tissue is often referred to asspongy or cancellous bone and is particularly concentrated near the endsof the bone (near the joints) and in the vertebrae of the spine. Thetrabecular tissue is characterized by small osteoid structures whichinter-connect with each other, as well as the more solid and densecortical tissue which makes up the outer surface and central shaft ofthe bone. This inter-connected network of trabeculae gives lateralsupport to the outer cortical structure and is critical to thebio-mechanical strength of the overall structure. In post-menopausalosteoporosis, it is, primarily, the net resorption and loss of thetrabeculae which leads to the failure and fracture of bone. In light ofthe loss of the trabeculae in post-menopausal women, it is notsurprising that the most common fractures are those associated withbones which are highly dependent on trabecular support, e.g., thevertebrae, the neck of the weight bearing bones such as the femur andthe fore-arm. Indeed, hip fracture, collies fractures, and vertebralcrush fractures are hall-marks of post-menopausal osteoporosis.

At this time, the only generally accepted method for treatment ofpost-menopausal osteoporosis is estrogen replacement therapy. Althoughtherapy is generally successful, patient compliance with the therapy islow primarily because estrogen treatment frequently produces undesirableside effects.

Throughout premenopausal time, most women have less incidence ofcardiovascular disease than age-matched men. Following menopause,however, the rate of cardiovascular disease in women slowly increases tomatch the rate seen in men. This loss of protection has been linked tothe loss of estrogen and, in particular, to the loss of estrogen'sability to regulate the levels of serum lipids. The nature of estrogen'sability to regulate serum lipids is not well understood, but evidence todate indicates that estrogen can upregulate the low density lipid (LDL)receptors in the liver to remove excess cholesterol. Additionally,estrogen appears to have some effect on the biosynthesis of cholesterol,and other beneficial effects on cardiovascular health.

It has been reported in the literature that post-menopausal women havingestrogen replacement therapy have a return of serum lipid levels toconcentrations to those of the pre-menopausal state. Thus, estrogenwould appear to be a reasonable treatment for this condition. However,the side-effects of estrogen replacement therapy are not acceptable tomany women, thus limiting the use of this therapy. An ideal therapy forthis condition would be an agent which would regulate the serum lipidlevel as does estrogen, but would be devoid of the side-effects andrisks associated with estrogen therapy.

The third major pathology associated with post-menopausal syndrome isestrogen-dependent breast cancer and, to a lesser extent,estrogen-dependent cancers of other organs, particularly the uterus.Although such neoplasms are not solely limited to a post-menopausalwomen, they are more prevalent in the older, post-menopausal population.Current chemotherapy of these cancers has relied heavily on the use ofanti-estrogen compounds such as, for example, tamoxifen. Although suchmixed agonist-antagonists have beneficial effects in the treatment ofthese cancers, and the estrogenic side-effects are tolerable in acutelife-threatening situations, they are not ideal. For example, theseagents may have stimulatory effects on certain cancer cell populationsin the uterus due to their estrogenic (agonist) properties and they may,therefore, be contraproductive in some cases. A better therapy for thetreatment of these cancers would be an agent which is an anti-estrogencompound having negligible or no estrogen agonist properties onreproductive tissues.

In response to the clear need for new pharmaceutical agents which arecapable of alleviating the symptoms of, inter alia, post-menopausalsyndrome, the present invention provides new benzothiophene compounds,pharmaceutical compositions thereof, and methods of using such compoundsfor the treatment of post-menopausal syndrome and other estrogen-relatedpathological conditions such as those mentioned below.

Uterine fibrosis (uterine fibroid disease) is an old and ever presentclinical problem which goes under a variety of names, including uterinefibroid disease, uterine hypertrophy, uterine lieomyomata, myometrialhypertrophy, fibrosis uteri, and fibrotic metritis. Essentially, uterinefibrosis is a condition where there is an inappropriate deposition offibroid tissue on the wall of the uterus.

This condition is a cause of dysmenorrhea and infertility in women. Theexact cause of this condition is poorly understood but evidence suggeststhat it is an inappropriate response of fibroid tissue to estrogen. Sucha condition has been produced in rabbits by daily administrations ofestrogen for 3 months. In guinea pigs, the condition has been producedby daily administration of estrogen for four months. Further, in rats,estrogen causes similar hypertrophy.

The most common treatment of uterine fibrosis involves surgicalprocedures both costly and sometimes a source of complications such asthe formation of abdominal adhesions and infections. In some patients,initial surgery is only a temporary treatment and the fibroids regrow.In those cases a hysterectomy is performed which effectively ends thefibroids but also the reproductive life of the patient. Also,gonadotropin releasing hormone antagonists may be administered, yettheir use is tempered by the fact they can lead to osteoporosis. Thus,there exists a need for new methods for treating uterine fibrosis, andthe methods of the present invention satisfy that need.

Endometriosis is a condition of severe dysmenorrhea, which isaccompanied by severe pain, bleeding into the endometrial masses orperitoneal cavity and often leads to infertility. The cause of thesymptoms of this condition appear to be ectopic endometrial growthswhich respond inappropriately to normal hormonal control and are locatedin inappropriate tissues. Because of the inappropriate locations forendometrial growth, the tissue seems to initiate local inflammatory-likeresponses causing macrophage infiltration and a cascade of eventsleading to initiation of the painful response. The exact etiology ofthis disease is not well understood and its treatment by hormonaltherapy is diverse, poorly defined, and marked by numerous unwanted andperhaps dangerous side effects.

One of the treatments for this disease is the use of low dose estrogento suppress endometrial growth through a negative feedback effect oncentral gonadotropin release and subsequent ovarian production ofestrogen; however, it is sometimes necessary to use continuous estrogento control the symptoms. This use of estrogen can often lead toundesirable side effects and even the risk of endometrial cancer.

Another treatment consists of continuous administration of progestinswhich induces amenorrhea and by suppressing ovarian estrogen productioncan cause regressions of the endometrial growths. The use of chronicprogestin therapy is often accompanied by the unpleasant CNS sideeffects of progestins and often leads to infertility due to suppressionof ovarian function.

A third treatment consists of the administration of weak androgens,which are effective in controlling the endometriosis; however, theyinduce severe masculinizing effects. Several of these treatments forendometriosis have also been implicated in causing a mild degree of boneloss with continued therapy. Therefore, new methods of treatingendometriosis are desirable.

Smooth aortal muscle cell proliferation plays an important role indiseases such as atherosclerosis and restenosis. Vascular restenosisafter percutaneous transluminal coronary angioplasty (PTCA) has beenshown to be a tissue response characterized by an early and late phase.The early phase occurring hours to days after PTCA is due to thrombosiswith some vasospasms while the late phase appears to be dominated byexcessive proliferation and migration of aortal smooth muscle cells. Inthis disease, the increased cell motility and colonization by suchmuscle cells and macrophages contribute significantly to thepathogenesis of the disease. The excessive proliferation and migrationof vascular aortal smooth muscle cells may be the primary mechanism tothe reocclusion of coronary arteries following PTCA, atherectomy, laserangioplasty and arterial bypass graft surgery. See "IntimalProliferation of Smooth Muscle Cells as an Explanation for RecurrentCoronary Artery Stenosis after Percutaneous Transluminal CoronaryAngioplasty," Austin et al., Journal of the American College ofCardiology, 8: 369-375 (Aug. 1985).

Vascular restenosis remains a major long term complication followingsurgical intervention of blocked arteries by percutaneous transluminalcoronary angioplasty (PTCA), atherectomy, laser angioplasty and arterialbypass graft surgery. In about 35% of the patients who undergo PTCA,reocclusion occurs within three to six months after the procedure. Thecurrent strategies for treating vascular restenosis include mechanicalintervention by devices such as stents or pharmacologic therapiesincluding heparin, low molecular weight heparin, coumarin, aspirin, fishoil, calcium antagonist, steroids, and prostacyclin. These strategieshave failed to curb the reocclusion rate and have been ineffective forthe treatment and prevention of vascular restenosis. See "Prevention ofRestenosis after Percutaneous Transluminal Coronary Angioplasty: TheSearch for a `Magic Bullet`," Hermans et al., American Heart Journal,122: 171-187 (July 1991).

In the pathogenesis of restenosis excessive cell proliferation andmigration occurs as a result of growth factors produced by cellularconstituents in the blood and the damaged arterial vessel wall whichmediate the proliferation of smooth muscle cells in vascular restenosis.

Agents that inhibit the proliferation and/or migration of smooth aortalmuscle cells are useful in the treatment and prevention of restenosis.The present invention provides for the use of compounds as smooth aortalmuscle cell proliferation inhibitors and, thus inhibitors of restenosis.

SUMMARY OF THE INVENTION

The present invention relates to compounds of formula I ##STR2## whereinR¹ is --H, --OH, --O(C₁ -C₄ alkyl), --OCOC₆ H₅, --OCO(C₁ -C₆ alkyl), or--OSO₂ (C₂ -C₆ alkyl);

R² is --H, --OH, --O(C₁ -C₄ alkyl), --OCOC₆ H₅, --OCO(C₁ -C₆ alkyl),--OSO₂ (C₂ -C₆ alkyl) or halo, providing when Z is --S--, R² is nothalo;

R³ is 1-piperidinyl, 1-pyrrolidinyl, methyl-1-pyrrolidinyl,dimethyl-1-pyrrolidinyl, 4-morpholino, dimethylamino, diethylamino,diisopropylamino, or 1-hexamethyleneimino;

n is 2 or 3; and

z is --O-- or --S--;

or a pharmaceutically acceptable salt thereof.

Also provided by the present invention are intermediate compounds offormula II which are useful for preparing the pharmaceutically activecompounds of the present invention, and are shown below ##STR3## whereinR^(1a) is --H or --OR⁷ in which R⁷ is a hydroxy protecting group;

R^(2a) is --H, halo, or --OR⁸ in which R⁸ is a hydroxy protecting group,providing when Z is --S--, R^(2a) is not halo;

R⁶ is --H or a hydroxy protecting group which can be selectivelyremoved; and

Z is --O-- or --S--.

The present invention further relates to pharmaceutical compositionscontaining compounds of formula I, optionally containing estrogen orprogestin, and the use of such compounds, alone, or in combination withestrogen or progestin, for alleviating the symptoms of post-menopausalsyndrome, particularly osteoporosis, cardiovascular related pathologicalconditions, and estrogen-dependent cancer. As used herein, the term"estrogen" includes steroidal compounds having estrogenic activity suchas, for example, 17β-estradiol, estrone, conjugated estrogen(Premarin®), equine estrogen 17β-ethynyl estradiol, and the like. Asused herein, the term "progestin" includes compounds havingprogestational activity such as, for example, progesterone,norethylnodrel, nongestrel, megestrol acetate, norethindrone, and thelike.

The compounds of the present invention also are useful for inhibitinguterine fibroid disease and endometriosis in women, and aortal smoothmuscle cell proliferation, particularly restenosis, in humans.

DETAILED DESCRIPTION OF THE INVENTION

One aspect of the present invention includes compounds of formula I##STR4## wherein R¹ is --H, --OH, --O(C₁ -C₄ alkyl), --OCOC₆ H₅,--OCO(C₁ -C₆ alkyl), or --OSO₂ (C₂ -C₆ alkyl);

R² is --H, --OH, --O(C₁ -C₄ alkyl), --OCOC₆ H₅, --OCO(C₁ -C₆ alkyl),--OSO₂ (C₂ -C₆ alkyl), or halo, providing when Z is --S--, R² is nothalo;

R³ is 1-piperidinyl, 1-pyrrolidinyl, methyl-1-pyrrolidinyl,dimethyl-1-pyrrolidinyl, 4-morpholino, dimethylamino, diethylamino,diisopropylamino, or 1-hexamethyleneimino;

n is 2 or 3; and

z is --O-- or --S--;

or a pharmaceutically acceptable salt thereof.

General terms used in the description of compounds herein described beartheir usual meanings. For example, "C₁ -C₆ alkyl" refers to straight orbranched aliphatic chains of 1 to 6 carbon atoms including moieties suchas methyl, ethyl, propyl, isopropyl, butyl, n-butyl, pentyl, isopentyl,hexyl, isohexyl, and the like. Similarly, the term "C₁ -C₄ alkoxy"represents a C₁ -C₄ alkyl group attached through an oxygen molecule andinclude moieties such as, for example, methoxy, ethoxy, n-propoxy,isopropoxy, and the like. Of these alkoxy groups, methoxy is highlypreferred in most circumstances.

The starting material for one route for preparing compounds of formula Iof the present invention, compounds of formula III, are preparedessentially as described by C. D. Jones in U.S. Pat. Nos. 4,418,068, and4,133,814, each of which is herein incorporated by reference. FormulaIII has the structure ##STR5## wherein R⁷ and R^(2a) are as definedabove.

The R⁷ and R⁸ hydroxy protecting groups are moieties which generally arenot found in the final, therapeutically active compounds of formula I,but which are intentionally introduced during a portion of the syntheticprocess to protect a group which otherwise might react in the course ofchemical manipulations, and is then removed at a later stage of thesynthesis. Since compounds bearing such protecting groups are ofimportance primarily as chemical intermediates (although somederivatives also exhibit biological activity), their precise structureis not critical. Numerous reactions for the formation, removal, andpossibly, reformation of such protecting groups are described in anumber of standard works including, for example, Protective Groups inOrganic Chemistry, Plenum Press (London and New York, 1973); Green, T.W., Protective Groups in Organic Synthesis, Wiley (New York, 1981); andThe Peptides, Vol. I, Schrooder and Lubke, Academic Press, (London andNew York, 1965).

Representative hydroxy protecting groups include, for example, -C₁ -C₄alkyl, -C₁ -C₄ alkoxy, --CO--(C₁ -C₆ alkyl), --SO₂ -- (C₄ -C₆ alkyl),and --CO--Ar in which Ar is optionally substituted phenyl. The term"substituted phenyl" refers to a phenyl group having one or moresubstituents selected from the group consisting of C₁ -C₄ alkyl, C₁ -C₄alkoxy, hydroxy, nitro, halo, and tri(chloro or fluoro) methyl. The term"halo" refers to bromo, chloro, fluoro, and iodo.

For compounds of formula III, preferred R⁷ and R⁸ substituents aremethyl and methoxymethyl. Compounds in which R⁷ and R⁸ each are methylare prepared via the procedure described in the above-referenced Jonespatent. Another preferred hydroxy protecting group is methoxymethyl.However, a formula IV compound, as shown below, is first preparedbearing the preferred methyl or other hydroxy protecting group(s). Theseprotecting groups are then removed, forming phenolic moieties, which arethen reprotected with methoxymethyl protecting groups.

The first steps of the present process for preparing certain compoundsof formula I include selectively placing a leaving group at the 3position of a formula III compound, coupling the reaction product of thefirst step with a 4-(protected-hydroxy)phenol, and removing the phenol'shydroxy protecting group. The present process is depicted in Scheme Ibelow. ##STR6##

In the first step of Scheme I, an appropriate leaving group isselectively placed at the 3-position of the formula III startingmaterial via standard procedures. Appropriate R⁹ leaving groups includethe sulfonates such as methanesulfonate, 4-bromobenzenesulfonate,toluenesulfonate, ethanesulfonate, isopropanesulfonate,4-methoxybenzenesulfonate, 4-nitrobenzenesulfonate,2-chlorobenzenesulfonate, triflate, and the like, halogens such asbromo, chloro, and iodo, and other related leaving groups. However, toinsure proper placement of the leaving group, the named halogens arepreferred, and bromo is especially preferred.

The present reaction is carried out using standard procedures. Forexample, when the preferred halogenating agents are used, an equivalentof such a halogenating agent, preferably bromine, is reacted with anequivalent of the formula III substrate, in the presence of a suitablesolvent such as, for example, chloroform or acetic acid. The reaction isrun at a temperature from about 40° C. to about 80° C.

The reaction product from the above process step, a compound of formulaIV, is then reacted with a 4-(protected-hydroxy)phenol to form compoundsof formula IIa in which R⁶ is a selectively removable hydroxy protectinggroup. Generally, the 4-hydroxy protecting moiety of the phenol may beany known protecting group which can be selectively removed withoutremoving the R⁷ and, when present, R⁸ moieties of a formula IIacompound. Preferred R⁶ protecting groups include methoxymethyl, when R⁷and/or R⁸ are not methoxymethyl, and benzyl. Of these, benzyl isespecially preferred. The 4-(protected-hydroxy)phenol reactants arecommercially available or can be prepared via standard procedures.

This coupling reaction is known in the art as an Ullman reaction and isrun according to standard procedures [see, e.g., Advanced OrganicChemistry: Reactions, Mechanisms, and Structure, Fourth Edition, 3-16,(J. March, ed., John Wiley & Sons, Inc. 1992); Jones, C. D., J. Chem.Soc. Perk. Trans. I, 4:407 (1992)].

In general, equivalent amounts of the two aryl substrates, in thepresence of up to an equimolar amount of a copper(I) oxide catalyst andan appropriate solvent, are heated to reflux under an inert atmosphere.Preferably, an equivalent of a formula IV compound in which R⁹ is bromois reacted with an equivalent amount of 4-benzyloxyphenol in thepresence of an equivalent of cuprous oxide.

Appropriate solvents for this reaction are those solvents or mixture ofsolvents which remain inert throughout the reaction. Typically, organicbases, particularly a hindered base such as, for example,2,4,6-collidine, are preferred solvents.

The temperature employed in this step should be sufficient to effectcompletion of this coupling reaction, and will influence the amount oftime required therefor. When the reaction mixture is heated to refluxunder an inert atmosphere such as nitrogen, the time-to-completionusually will be from about 20 to about 60 hours.

Following coupling, which forms a formula IIa compound, formula IIbcompounds are prepared by selectively removing the R⁶ hydroxy protectinggroup of a formula IIa compound via well known reduction procedures. Itis imperative that the selected procedure will not affect the R⁷ and,when present, R⁸ hydroxy protecting groups.

When R⁶ is the preferred benzyl moiety, and R⁷ and, when present, R⁸each are methyl, the present process step is carried out via standardhydrogenolysis procedures. Typically, the formula IIa substrate is addedto a suitable solvent or mixture of solvents, followed by the additionof a proton donor to accelerate the reaction and an appropriatehydrogenation catalyst.

Appropriate catalysts include noble metals and oxides such as palladium,platinum, and rhodium oxide on a support such as carbon or calciumcarbonate. Of these, palladium-on-carbon, particularly 10%palladium-on-carbon, is preferred.

Solvents for this reaction are those solvents or mixture of solventswhich remain inert throughout the reaction. Typically, ethylacetate andC₁ -C₄ aliphatic alcohols, particularly ethanol, is preferred.

For the present reaction, hydrochloric acid serves as an adequate andpreferred proton donor.

When run at ambient temperature and a pressure ranging form about 30 psito about 50 psi, the present reaction runs quite rapidly. Progress ofthis reaction may be monitored by standard chromatographic techniquessuch as thin layer chromatography.

Compounds of formula IIa and IIb are novel, are encompassed within thegenus described herein as formula II compounds, and are useful forpreparing the pharmaceutically active compounds of formula I.

Upon preparation of a formula IIb compound, it is reacted with acompound of formula V

    R.sup.3 --(CH.sup.2).sub.n --Q                             V

wherein R³ and n are as defined above, and Q is a bromo or, preferably,a chloro moiety, to form a compound of formula VI. The formula VIcompound is then deprotected to form a compound of formula Ia. Thesesteps of the present process are shown in Scheme II below ##STR7##wherein R³, R⁷, R^(2a), and n are as defined above, and R^(2b) is --H,--OH, or halo.

In the first step of the process shown in Scheme II, the alkylation iscarried out via standard procedures. Compounds of formula V arecommercially available or are prepared by means well known to one ofordinary skill in the art. Preferably, the hydrochloride salt of aformula V compound, particularly 2-chloroethylpiperidine hydrochloride,is used.

Generally, at least about 1 equivalent of formula IIb substrate arereacted with 2 equivalents of a formula V compound in the presence of atleast about 4 equivalents of an alkali metal carbonate, preferablycesium carbonate, and an appropriate solvent.

Solvents for this reaction are those solvents or mixture of solventswhich remain inert throughout the reaction. N,N-dimethylformamide,especially the anhydrous form thereof, is preferred.

The temperature employed in this step should be sufficient to effectcompletion of this alkylation reaction. Typically, ambient temperatureis sufficient and preferred.

The present reaction preferably is run under an inert atmosphere,particularly nitrogen.

Under the preferred reaction conditions, this reaction will run tocompletion in about 16 to about 20 hours. Of course, the progress of thereaction can be monitored via standard chromatographic techniques.

As an alternative for preparing compounds of formula VI, a formula IIbcompound is reacted with an excess of an alkylating agent of the formula

    Q--(CH.sub.2).sub.n --Q'

wherein Q and Q' each are the same or different leaving group, in analkali solution. Appropriate leaving groups are the aforementionedleaving groups used in the preparation of compounds of formula IV.

A preferred alkali solution for this alkylation reaction containspotassium carbonate in an inert solvent such as, for example, methyethylketone (MEK) or DMF. In this solution, the 4-hydroxy group of thebenzoyl moiety of a formula IIb compound exists as a phenoxide ion whichdisplaces one of the leaving groups of the alkylating agent.

This reaction is best when the alkali solution containing the reactantsand reagents is brought to reflux and allowed to run to completion. Whenusing MEK as the preferred solvent, reaction times run from about 6hours to about 20 hours.

The reaction product from this step is then reacted with 1-piperidine,1-pyrrolidine, methyl-1-pyrrolidine, dimethyl-1-pyrrolidine,4-morpholine, dimethylamine, diethylamine, diisopropylamine, or1-hexamethyleneimine, via standard techniques, to form compounds offormula VI. Preferably, the hydrochloride salt of piperidine is reactedwith the alkylated compound of formula IIb in an inert solvent, such asanhydrous DMF, and heated to a temperature in the range from about 60°C. to about 110° C. When the mixture is heated to a preferredtemperature of about 90° C., the reaction only takes about 30 minutes toabout 1 hour. However, changes in the reaction conditions will influencethe amount of time this reaction needs to be run for completion. Ofcourse, the progress of this reaction step can be monitored via standardchromatographic techniques.

Compounds of formula VI, in which R⁷ and when present, R⁸ each are C₁-C₄ alkyl, preferably methyl, and in which R^(2a) is --H or halo, arenovel and are pharmaceutically active for the methods herein described.Accordingly, such compounds are encompassed by the definition herein ofcompounds of formula I.

Preferred compounds of formula I are obtained by cleaving the R⁷ and,when present, R⁸ hydroxy protecting groups of formula VI compounds viawell known procedures. Numerous reactions for the formation and removalof such protecting groups are described in a number of standard worksincluding, for example, Protective Groups in Organic Chemistry, PlenumPress (London and New York, 1973); Green, T. W., Protective Groups inOrganic Synthesis, Wiley, (New York, 1981); and The Peptides, Vol. I,Schrooder and Lubke, Academic Press (London and New York, 1965). Methodsfor removing preferred R⁷ and/or R⁸ hydroxy protecting groups,particularly methyl and methoxymethyl, are essentially as described inthe Examples, infra.

Compounds of formula Ia are novel, are pharmaceutically active for themethods herein described, and are encompassed by formula I as definedherein.

Compounds of formula I in which R¹ is --H are prepared via the syntheticroute shown below in Scheme III. Using this route, a 3-position leavinggroup (R⁹) is placed on commercially available thianaphthene (formulaVII) to form a compound of formula VIII, which is then coupled with a4-(protected-hydroxy)phenol, providing compounds of formula IX. ##STR8##wherein R⁶ is a hydroxy protecting group which can be selectivelyremoved and R⁹ is a leaving group.

The compound of formula VII is commercially available. Preparation offormulae VIII and IX compounds, including the definition of R⁶ and R⁹substituents, as well as preferred reactants and conditions, unlessotherwise herein stated, are the same as described above and shown inScheme I, supra.

Compounds of formula IX are then arylated via Suzuki coupling [see,e.g., Suzuki, A., Pure and Appl. Chem., 6(2):213-222 (1994)]. Using oneSuzuki coupling option, a formula IX compound is selectively halogenatedat the 2-position, and then coupled with an arylboronic acid compound offormula XIa (Route A).

Preferably, however, an arylboronic acid of formula Xb is formed from acompound of formula IX, and then reacted with a halo-arene of formulaXIb to give novel intermediates of formula IIc (Route B). Such novelintermediates are useful for preparing pharmaceutically active compoundsof the present invention (formula Ib compounds) via alkylation anddeprotection. These reactions are shown below in Scheme IV. ##STR9##wherein R², R^(2b), R³, R⁶ and n are as defined above;

X is iodo, bromo, or fluoro, in the order of preference; and

X' is iodo, bromo, or fluoro, in the order of preference, or triflate.

The first step in Route A in Scheme IV is the 2-position iodination orbromination of a formula IX compound using standard procedures.Generally, a formula IX compound is reacted with a slight excess ofn-butyllithium in hexane, in an appropriate solvent and under an inertatmosphere such as nitrogen, followed by the dropwise addition of aslight excess of the desired halogenating agent in an appropriatesolvent. Preferably, the halogenating agent for this step is iodine, butthe use of bromine, N-bromosuccinimide is also permitted.

Appropriate solvents include an inert solvent or mixture of solventssuch as, for example, diethyl ether, dioxane, and tetrahydrofuran (THF).Of these, tetrahydrofuran, particularly anhydrous THF, is preferred.

The present selective, 2-position halogenation reaction optionally isrun at a temperature from about -75° C. to about 85° C.

The product of the above reaction, a halo-arene of formula Xa, is thencoupled with an arylboronic acid of formula XIa, via standard Suzukicoupling procedures, to provide compounds of formula IIc. Compounds offormula XIa, in which R^(2a) is --H, halo, or --OR⁸ (R⁸ is a hydroxyprotecting group as defined, supra) are derived from commerciallyavailable compounds via procedures well known to one of ordinary skillin the art (See, e.g., March J.; and Suzuki, A., supra).

In the present coupling reaction, a slight excess of a formula XIacompound is reacted with each equivalent of a formula Xa compound in thepresence of a palladium catalyst and an appropriate base in an inertsolvent such as toluene.

Although various palladium catalysts drive Suzuki coupling reactions,the catalyst selected usually is reaction specific. Thus, the use oftetrakis triphenylphosphine palladium in the present reaction is highlypreferred.

Likewise, various bases may be used in the present coupling reaction.However, it is preferred to use an alkali metal carbonate, particularly2N sodium carbonate.

The temperature employed in this step should be sufficient to effectcompletion of the coupling reaction. Typically, heating the reactionmixture to reflux for a period from about 2 to about 4 hours is adequateand preferred.

In Route B of Scheme IV, a 2-position arylboronic of formula Xb isprepared using well known procedures. Generally, a compound of formulaIX is treated with a slight excess of n-butyllithium in hexanes, in anappropriate solvent and under an inert atmosphere such as nitrogen,following by the dropwise addition of an appropriate trialkylborate.

Appropriate solvents include an inert solvent or mixture of solventssuch as, for example, diethyl ether, dioxane, and tetrahydrofuran (THF).THF, particularly anhydrous THF, is preferred.

The preferred trialkylborate used in the present reaction istriisopropyl borate.

The product of this reaction, a compound of formula Xb, is then reactedwith a aryl halide or aryl triflate of formula XIb, via standard Suzukicoupling procedures, to provide compounds of formula IIc. The preferredreaction conditions for the present reaction are as described for thereaction of compounds of formulae XIa and Xa, in Scheme IV, which alsoprovide compounds of formula IIc.

The transformation of compounds of formula IIc to formula Ia compoundsis carried out as described above for the conversion of formula IIacompounds to compounds of formula Ia.

Compounds of formulae IIc and IId are novel, and are useful for thepreparation of pharmaceutically active compounds of the presentinvention.

Compounds of formulae XII and Ib also are novel, are useful for themethods herein described, and are encompassed by formula I as hereindefined.

As an alternative to the preparation of formula Ib compounds as shown inScheme IV, supra, such compounds can be prepared starting from acompound of formula XIII, and using the synthetic sequence as shown inScheme I, supra, and the associated above described preferredexperimental conditions.

Compounds of formula XIII ##STR10## wherein R^(2a) is as defined above,are known in the art and are described by Jones, C. D., et al., in theafore-mentioned U.S. Pat. Nos. 4,133,814 and 4,418,068.

For preparation of compounds of formula I in which Z is S, a compound offormula IV above is metallated, the resulting product is reacted with a4-(protected-hydroxy)phenyl disulfide, and the phenol protecting groupis selectively removed. This general process is shown below in Scheme V.##STR11## wherein R^(1a) is --H or --OR⁷ and R⁷ is a hydroxy protectinggroup;

R^(2a) is --H, or --OR⁸ and R⁸ is a hydroxy protecting group;

R⁶ is a hydroxy protecting group which can be selectively removed;

R⁹ is a leaving group; and

M is a metal ion.

In the first two steps of Scheme V, a formula IVa compound is metallatedvia well known procedures. Most commonly, and preferably, a formula IVacompound is treated with a slight excess of n-butyllithium in hexanes inan appropriate solvent, followed by the dropwise addition of a solutionof a disulfide compound of formula XV in an appropriate solvent.

Both of these reaction steps are run under an inert atmosphere such asnitrogen, while appropriate solvents for both steps include one or moreinert solvents such as diethyl ether, dioxane, and THF. Of these, THF,particularly the anhydrous form thereof, is preferred. In addition, thepresent reaction steps are run at a temperature from about -78° C. toabout 85° C.

In the first step of the present reaction, a metallated compound offormula XIV is provided. The 4-(protected-hydroxy)phenyl disulfide (aformula XV compound) which is reacted with such a formula XIV compoundto give a compound of formula IIe, is prepared by protection of thehydroxy group of commercially available 4-hydrogenphenylsulfide with anappropriate protecting group according to conditions known in the art. Apreferred R⁶ protecting group is methoxymethyl, providing R⁷ and R⁸, ifeither or both are present, is a hydroxy protecting group other thanmethoxymethyl. It is imperative that the R⁶ hydroxy protecting group isa moiety different than those formed by R⁷ and R⁸ hydroxy protectinggroups, when present, so that the R⁶ group can selectively be removed,via standard procedures, to provide compounds of formula IIf.

To effect deprotection by removal of the R⁶ protecting group, a formulaIIe compound in a protic solvent or mixture of solvents is reacted in anacid media containing at least one equivalent of acid, preferablymethanesulfonic acid, and heating from about 25° to about 110° C.Typically, the reaction time is from about 6 to about 24 hours, but theprogress of the reaction may be monitored via standard chromotographictechniques.

Appropriate solvents for the present reaction include, for example,water and methanol.

Compounds of formulae IIe and IIf are novel, are useful for preparingpharmaceutically active compounds of formula I and are hereinencompassed within the above depiction of formula II.

Compounds of formula Ic ##STR12## wherein R^(1b) is --H or --OH;

R^(2b) is --H or --OH; and

R³ and n, are as defined above, are prepared by using theabove-described procedures related to the process steps shown in SchemesII and IV. Such compounds of formula Ic also are novel, are useful forthe methods of the present invention, and are herein encompassed withinthe above depiction of formula I.

As an alternative, compounds of formula I on which either R¹ or R² is--H and the other R¹ or R² substituent is --OH can be prepared fromcompounds of formula I in which both R¹ or R² are --OH. The dihydroxycompound of formula I is converted to a mixture of 6- and4'-monotriflates, and the triflate moiety is reduced to hydrogen [see,Saa, J. M., et al., J. Org. Chem., 55:991 (1990)]. The resulting mixtureof monohydroxy derivatives, either as the free base or pharmaceuticallyacceptable salt, preferrably the hydrochloride salt, can then beseparated by standard crystallization techniques.

In general, a dihydroxy compound of formula I is treated with about 4 toabout 6 equivalents of a amine base, such as triethylamine, in anon-reactive solvent followed by the addition of 1 equivalent oftrifluoromethanesulfonic anhydride. A statistical mixture of mono- andbitriflates are produced and separated by standard chromatographictechniques. A preferred solvent for this step is anhydrousdichloromethane.

When run at a temperature range from about 0° C. to about 25° C., thepresent reaction is complete within from about 1 to about 5 hours.

The isolated mixture of monotriflated compounds is then hydrogenated, ina non-reactive solvent, in the presence of from about 3 to about 6equivalents of a amine base, preferably triethylamine, and ahydrogenation catalyst such as palladium on carbon which is preferred.Preferred solvents for this reaction include ethyl acetate and ethanolor, alternatively, a mixture thereof. When this step of the presentreaction is run under about 40 psi of H₂, at ambient temperature, thereaction time is from about 2 to about 5 hours.

The resulting mixture of nonhydroxy derivatives of formula I havedifferent solubilities in ethyl aceate and the 6-hydroxy-4'-hydrogenderivatives can be partially separated from the 6-hydrogen-4'-hydroxyderivatives by selective crystallization. Further separation, whichprovides pure monohydroxy compounds of formula I, can be achieved byconversion of the enriched mixtures to the hydrochloride salts followedby crystallization from ethyl acetate-ethanol.

Other preferred compounds of formula I are prepared by replacing 6-and/or 4'-position hydroxy moieties, when present, with a moiety of theformula --O--CO--(C₁ -C₆ alkyl), or --O--SO₂ --(C₂ -C₆ alkyl) via wellknown procedures. See, e.g., U.S. Pat. No. 4,358,593.

For example, when an --O--CO(C₁ -C₆ alkyl) group is desired, a mono- ordihydroxy compound of formula I is reacted with an agent such as acylchloride, bromide, cyanide, or azide, or with an appropriate anhydrideor mixed anhydride. The reactions are conveniently carried out in abasic solvent such as pyridine, lutidine, quinoline or isoquinoline, orin a tertiary amine solvent such as triethylamine, tributylamine,methylpiperidine, and the like. The reaction also may be carried out inan inert solvent such as ethyl acetate, dimethylformamide,dimethylsulfoxide, dioxane, dimethoxyethane, acetonitrile, acetone,methyl ethyl ketone, and the like, to which at least one equivalent ofan acid scavenger (except as noted below), such as a tertiary amine, hasbeen added. If desired, acylation catalysts such as4-dimethylaminopyridine or 4-pyrrolidinopyridine may be used. See, e.g.,Haslam, et al., Tetrahedron, 36:2409-2433 (1980).

The present reactions are carried out at moderate temperatures, in therange from about -25° C. to about 100° C., frequently under an inertatmosphere such as nitrogen gas. However, ambient temperature is usuallyadequate for the reaction to run.

Acylation of a 6-position and/or 4'-position hydroxy group also may beperformed by acid-catalyzed reactions of the appropriate carboxylicacids in inert organic solvents. Acid catalysts such as sulfuric acid,polyphosphoric acid, methanesulfonic acid, and the like are used.

The aforementioned R¹ and/or R² groups of formula I compounds also maybe provided by forming an active ester of the appropriate acid, such asthe esters formed by such known reagents such asdicyclohexylcarbodiimide, acylimidazoles, nitrophenols,pentachlorophenol, N-hydroxysuccinimide, and 1-hydroxybenzotriazole.See, e.g., Bull. Chem. Soc. Japan, 38:1979 (1965), and Chem. Ber., 788and 2024 (1970).

Each of the above techniques which provide --O--CO--(C₁ -C₆ alkyl)moieties are carried out in solvents as discussed above. Thosetechniques which do not produce an acid product in the course of thereaction, of course, do not call for the use of an acid scavenger in thereaction mixture.

When a formula I compound is desired in which the 6- and/or 4'-positionhydroxy group of a formula I compound is converted to a group of theformula --O--SO₂ --(C₂ -C₆ alkyl), the mono- or dihydroxy compound isreacted with, for example, a sulfonic anhydride or a derivative of theappropriate sulfonic acid such as a sulfonyl chloride, bromide, orsulfonyl ammonium salt, as taught by King and Monoir, J. Am. Chem. Soc.,97:2566-2567 (1975). The dihydroxy compound also can be reacted with theappropriate sulfonic anhydride or mixed sulfonic anhydrides. Suchreactions are carried out under conditions such as were explained abovein the discussion of reaction with acid halides and the like.

Although the free-base form of formula I compounds can be used in themethods of the present invention, it is preferred to prepare and use apharmaceutically acceptable salt form. Thus, the compounds used in themethods of this invention primarily form pharmaceutically acceptableacid addition salts with a wide variety of organic and inorganic acids,and include the physiologically acceptable salts which are often used inpharmaceutical chemistry. Such salts are also part of this invention.Typical inorganic acids used to form such salts include hydrochloric,hydrobromic, hydroiodic, nitric, sulfuric, phosphoric, hypophosphoric,and the like. Salts derived from organic acids, such as aliphatic monoand dicarboxylic acids, phenyl substituted alkanoic acids,hydroxyalkanoic and hydroxyalkandioic acids, aromatic acids, aliphaticand aromatic sulfonic acids, may also be used. Such pharmaceuticallyacceptable salts thus include acetate, phenylacetate, trifluoroacetate,acrylate, ascorbate, benzoate, chlorobenzoate, dinitrobenzoate,hydroxybenzoate, methoxybenzoate, methylbenzoate, o-acetoxybenzoate,naphthalene-2-benzoate, bromide, isobutyrate, phenylbutyrate,β-hydroxybutyrate, butyne-1,4-dioate, hexyne-1,4-dioate, caprate,caprylate, chloride, cinnamate, citrate, formate, fumarate, glycollate,heptanoate, hippurate, lactate, malate, maleate, hydroxymaleate,malonate, mandelate, mesylate, nicotinate, isonicotinate, nitrate,oxalate, phthalate, terephthatate, phosphate, monohydrogenphosphate,dihydrogenphosphate, metaphosphate, pyrophosphate, propiolate,propionate, phenylpropionate, salicylate, sebacate, succinate, suberate,sulfate, bisulfate, pyrosulfate, sulfite, bisulfite, sulfonate,benzenesulfonate, p-bromophenylsulfonate, chlorobenzenesulfonate,ethanesulfonate, 2-hydroxyethanesulfonate, methanesulfonate,naphthalene-1-sulfonate, naphthalene-2-sulfonate, p-toluenesulfonate,xylenesulfonate, tartarate, and the like. Preferred salts are thehydrochloride and oxalate salts.

The pharmaceutically acceptable acid addition salts are typically formedby reacting a compound of formula I with an equimolar or excess amountof acid. The reactants are generally combined in a mutual solvent suchas diethyl ether or ethyl acetate. The salt normally precipitates out ofsolution within about one hour to 10 days and can be isolated byfiltration or the solvent can be stripped off by conventional means.

The pharmaceutically acceptable salts generally have enhanced solubilitycharacteristics compared to the compound from which they are derived,and thus are often more amenable to formulation as liquids or emulsions.

The following examples are presented to further illustrate thepreparation of compounds of the present invention. It is not intendedthat the invention be limited in scope by reason of any of the followingexamples.

NMR data for the following Examples were generated on a GE 300 MHz NMRinstrument, and anhydrous d-6 DMSO was used as the solvent unlessotherwise indicated.

PREPARATION 1 Preparation of[3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophene[3-(4-benzyloxy)phenoxy]benzo[b]thiophene ##STR13##

To a solution of 3-bromo-benzo[b]thiophene (69.62 g, 0.325 mol) in 55 mLof anhydrous collidine under N₂ was added 4-benzyloxyphenol (97.6 g,0.488 mol) and cuprous oxide (23.3 g, 0.163 mol). The mixture was heatedto reflux for 24 hours. Upon cooling, the reaction mixture was dilutedwith ethyl acetate (200 mL) and the crude mixture filtered through a padof Celite® (Aldrich, Milwaukee, Wis.) to remove inorganic salts. Thefiltrate was washed with 1N hydrochloric acid (3×150 mL). The organicwas dried (sodium sulfate) and concentrated in vacuo to a liquid.Thianaphthene was removed by distillation (10 mm Hg, 115°-120° C.). Theremainder of the material was chromatographed (silicon dioxide, hexanes:ethyl acetate 85:15) to provide 12.2 g of benzo[b]thiophene and 12.95 g(35% based on recovered starting material) of[3-(4-benzyloxy)phenoxy]benzo-[b]thiophene as an off-white solid. mp84°-86 ° C. ¹ H NMR (CDCl₃) δ 7.91-7.83 (m, 2H), 7.47-7.34 (m, 7H), 7.04(q, J_(AB) =9.0 Hz, 4H), 6.47 (s, 1H), 5.07 (s, 2H). Anal. Calcd. forC₂₁ H₁₆ O₂ S: C, 75.88; H, 4.85. Found: C, 75.75; H, 5.00.

PREPARATION 2 [2-Iodo-3-(4-benzyloxy)phenoxy]benzo-[b]thiophene##STR14##

To a solution of [3-(4-benzyloxy)phenoxy]benzo[b]thiophene (6.00 g, 18.1mmol) in anhydrous tetrahydrofuran (100 mL) under N₂ at -78° C. wasadded n-butyllithium (12.4 mL, 19.9 mmol, 1.6M in hexanes) dropwise viasyringe. The solution turned from colorless to deep orange. Afterstirring for 20 minutes at -78° C., the lithio species was treated withI₂ (5.03, 19.9 mmol), added dropwise via canula as a solution in 50 mLof anhydrous tetrahydrofuran. Upon completion of the addition, thereaction turned light yellow in color, and was allowed to slowly warm toroom temperature. The reaction was quenched by the addition of 0.1Nsodium sulfite solution (200 mL). The layers were separated and theaqueous extracted with ethyl acetate (2×150 mL). The organic wascombined, dried (sodium sulfate), and concentrated in vacuo to give anoil that crystallized on standing. Recrystallization from hexanes/ethylether yielded 7.10 g (86%) of [2-Iodo-3-(4-benzyloxy)phenoxy]benzo[b]thiophene as a white crystallinepowder. mp 87°-92° C. ¹ H NMR (CDCl₃) δ 7.72 (d, J=8.1 Hz, 1H),7.47-7.20 (m, 8H), 6.89 (s, 4H), 5.01 (s, 2H). Anal. Calcd. for C₂₁ H₁₅O₂ SI: C, 55.03; H, 3.30. Found: C, 55.29; H, 3.31.

PREPARATION 3[2-(4-tertbutyloxyphenyl)-3-(4-benzyloxy)phenoxy]benzo[b]thiophene##STR15##

To a solution of [2-Iodo-3-(4-benzyloxy)phenoxy]benzo[b]thiophene (4.50g, 9.82 mmol) in toluene (20 mL) was added 4-(tertbutoxy)phenyl boronicacid (2.28 g, 11.75 mmol) followed bytetrakistriphenylphosphinepalladium (0.76 g, 0.66 mmol). To thissolution was added 14.5 mL of 2N sodium carbonate solution. Theresulting mixture was heated to reflux for 3 hours. Upon cooling, thereaction was diluted with 150 mL of ethyl acetate. The organic waswashed with 0.1N sodium hydroxide (2×100 mL) and then dried (sodiumsulfate). Concentration produced a semi-solid that was dissolved inchloroform and passed through a pad of silicon dioxide. Concentrationproduced an oil that was triturated from hexanes to yield 4.00 g (91%)of [2-(4-tertbutyloxyphenyl)-3-(4-benzyloxy)phenoxy]benzo[b]thiophene asa white powder. mp 105°-108° C. ¹ H NMR (CDCl₃) δ 7.77 (d, J=7.7 Hz, 1H), 7.68 (d, J=8.6 Hz, 2H), 7.43-7.24 (m, 8H), 6.98 (d, J=8.6 Hz, 2H),6.89 (q, J_(AB) =9.3 Hz, 4H), 4.99 (s, 2H), 1.36 (s, 9H). FD mass spec:480. Anal. Calcd. for C₃₁ H₂₈ O₃ S: C, 77.47; H, 5.87. Found: C, 77.35;H, 5.99.

PREPARATION 4 Prepared in a Similar Manner Employing4-Methoxyphenylboronic Acid was[2-(4-methoxyphenyl)-3-(4-benzyloxy)phenoxy]benzo[b]-thiophene ##STR16##

Yield=73%. mp=115°-118° C. ¹ H NMR (CDCl₃) δ 7.80-7.90 (m, 3H),7.33-7.53 (m, 8H), 6.93-7.06 (m, 6H), 5.00 (s, 2H), 3.83 (s, 3H). FDmass spec: 438. Anal. Calcd. for C₂₈ H₂₂ O₃ S: C, 76.69; H, 5.06. Found:C, 76.52; H, 5.09.

PREPARATION 5[2-(4-tertbutyloxyphenyl)-3-(4-hydroxy)phenoxy]benzo[b]thiophene##STR17##

To a solution of[2-(4-tertbutyloxyphenyl)-3-(4-benzyloxy)phenoxy]benzo[b]thiophene (1.50g, 3.37 mmol) in 30 mL of absolute ethanol containing 1% concentratedhydrochloric acid was added 0.50 g of 10% palladium-on-carbon. Themixture was hydrogenated at 40 psi for 1 hour, after which the reactionwas judged to be complete by thin layer chromatography. The mixture wasfiltered through a pad of Celite, and the filtrate concentrated invacuo. The crude product was dissolved in minimal ethyl acetate andpassed through a short silicon dioxide column to remove Celite (ethylacetate as eluant). Concentration provided a white solid that wastriturated from hexanes/ethyl ether. Filtration provided 868 mg (73%) of[2-(4-tertbutyloxyphenyl)-3-(4-hydroxy)phenoxy]-benzo[b]thiophene. mp210°-213° C. ¹ H NMR (DMSO-d₆) δ 9.13 (s, 1H), 7.94 (d, J=7.7 Hz, 1H),7.63 (d, J=8.6 Hz, 2H), 7.35-7.26 (m, 3H), 7.01 (d, J=8.6 Hz, 2H), 6.70(q, J_(AB) =8.9 Hz, 4H), 1.28 (S, 9H). FD mass spec: 390. Anal. Calcd.for C₂₄ H₂₂ O₃ S: C, 73.82; H, 5.68. Found: C, 73.98; H, 5.84.

PREPARATION 4 Prepared in a Similar Manner was [2-(4-methoxyphenyl)-3-(4-hydroxy)phenoxy]benzo[b]thiophene ##STR18##

Yield=80%. mp=120°-125° C. ¹ H NMR (CDCl₃) δ 7.80-7.90 (m, 3H), 7.48 (m,1H), 7.30-7.48 (m, 2H), 6.90-7.03 (m, 4H), 6.76-6.86 (m, 2H), 3.82 (s,3H). FD mass spec: 348; Anal Calcd. for C₂₁ H₁₆ O₃ S: C, 72.39; H, 4.63.Found: C, 72.68; H, 4.82.

EXAMPLE 1[3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophene##STR19##

To a solution of [2-(4-tertbutyloxyphenyl)-3-(4-hydroxy)phenoxy]benzo[b]thiophene (1.25 g, 3.20 mmol) in anhydrousN,N-dimethylformamide (10 mL) at ambient temperature was added cesiumcarbonate (5.70 g, 17.6 mmol). After stirring for 20 minutes,2-chloroethylpiperidine hydrochloride (1.95 g, 10.56 mmol) was added insmall portions. The resulting heterogeneous mixture was stirredvigorously for 24 hours. The contents of the reaction were then dilutedwith water (200 mL). The aqueous phase was extracted with ethyl acetate(3×100 mL). The combined organic layer was then washed with water (2×200mL). Drying of the organic layer (sodium sulfate) and concentration invacuo gave an oil. Chromatography (5-10% methanol/chloroform) provided1.47 g (91%) of3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-tertbutyloxyphenyl)]benzo[b]-thiophenethat was carried on directly to the next step without characterization.

3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-tertbutyloxyphenyl)]benzo[b]thiophene(1.37 g, 2.73 mmol) was dissolved in triflouroacetic acid (10 mL) atambient temperature. After stirring for 15 minutes, the solvent wasremoved in vacuo. The residue was dissolved in ethyl acetate (20 mL) andwashed with sat. sodium bicarbonate solution (3×10 mL). The organiclayer was dried (sodium sulfate) and concentrated whereupon a whitesolid precipitated formed in solution. The product was recrystallizedfrom ethyl acetate-ethyl ether to provide 1.03 g (85%) of3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]-thiophene as colorless crystals. mp 169°-172°C. ¹ H NMR (DMSO-d₆) δ 9.81 (s, 1H), 7.93 (d, J=7.7 Hz, 1H), 7.54 (d,J=8.5 Hz, 2H), 7.36-7.26 (m, 3H), 6.86 (s, 4H), 6.78 (d, J=8.6 Hz, 2H),4.10 (m, 2H), 3.29 (m, 2H), 2.95-2.75 (m, 4H), 1.68-1.40 (m, 6H). Anal.Calcd. for C₂₇ H₂₇ NO₃ S.0.55 CF₃ CO₂ H: C, 66.40; H, 5.46; N, 2.76.Found: C, 65.99; H, 5.49; N, 2.61

EXAMPLE 23-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophenewas Converted to its Hydrochloride Salt in 90% Yield by Treatment withEthyl Ether.hydrochloric Acid in Ethyl Acetate ##STR20##

Data for Example 2 mp 233°-240° C. ¹ H NMR (DMSO-d₆) δ 10.43 (m, 1H),9.89 (s, 1H), 7.93-7.95 (m, 1H), 7.60-7.64 (m, 2H), 7.35-7.50 (m, 3H),6.83-7.03 (m, 6H), 4.27-4.30 (m, 2H), 3.40-3.60 (m, 4H), 2.96-3.10 (m,2H), 1.70-1.95 (m, 5H), 1.40-1.53 (m, 1H). FD mass spec: 446. Anal.Calcd. for C₂₇ H₂₇ NO₃ S.1.0HCl: C, 67.28; H, 5.86; N, 2.91. Found: C,67,07; H, 5.66; N, 2.96.

EXAMPLE 3

Prepared in an analogous manner were the following examples:

[3-[4-[2-(1-pyrolidinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophene##STR21##

mp 150°-155° C. ¹ H NMR (DMSO-d₆) δ 9.79 (s, 1H), 7.92 (d, J=7.8 Hz,1H), 7.54 (d, J=8.6 Hz, 2H), 7.36-7.26 (m, 3H), 6.84 (s, 4H), 6.78 )d,J=8.6 Hz, 2H), 4.00 (bt, 2H), 2.92 (m, 2H), 2.85 (m, 4H), 1.73 (m, 4H).Anal. Calcd. for C₂₆ H₂₅ NO₃ S.0.33 CF₃ CO₂ H: C, 68.25; H, 5.44; N,2.99. Found: C, 68.29; H, 5.46; N, 3.19.

EXAMPLE 4[3-[4-[2-(1-hexamethyleneimino)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophene##STR22##

mp 189°-191° C. ¹ H NMR (DMSO-d₆) δ 7.91 (d, J=7.6 Hz, 1H), 7.52 (d,J=8.5 Hz, 2H), 7.34-7.25 (m, 3H), 6.81 (s, 4H), 6.75 (d, J=8.6 HZ, 2H),3.89 (bt, 2H), 2.75 (bt, 2H), 2.68 (m, 4H), 1.48 (m, 8H). Anal. Calcd.for C₂₈ H₂₉ NO₃ S.1.50 H₂ O: C, 69.11; H, 6.79; N, 2.88. Found: C,69.25; H, 6.79; N, 2.58.

EXAMPLE 5[3-[4-[2-(1-N,N-diethylamino)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophene##STR23##

mp 70° C. ¹ H NMR (DMSO-d₆) δ 9.91 (bs, 1H), 7.92 (d, J=7.9 Hz, 1H),7.54 (d, J=8.6 Hz, 2H), 7.35-7.24 (m, 3H), 6.82 (s, 4H), 6.78 (d, J=8.6Hz, 2H), 3.88 (bt, 2H), 2.76 (bt, 2H), 2.51 (m, 4H), 0.91 (m, 6H). FDmass spec: 434. Anal. Calcd. for C₂₆ H₂₇ NO₃ S.0.50 H₂ O: C, 70.56; H,6.38; N, 3.16 Found: C, 70.45; H, 6.26; N, 3.20.

EXAMPLE 63-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-methoxyphenyl)]benzo[b]thiophenehydrochloride ##STR24##

mp=228°-230° C. ¹ H NMR (DMSO-d₆) δ 7.96 (d, J=7.5 Hz, 1H), 7.66 (d,J=8.8 Hz, 2H), 7.35-7.50 (m, 3H), 6.98 (d, J=8.7 Hz, 2H), 6.86-6.90 (m,4H), 4.28-4.31 (m, 2H), 3.74 (s, 3H), 3.37-3.45 (m, 4H), 2.92-2.96 (m,2H), 2.46-2.48 (m, 5H), 1.74 (m, 1H). FD mass spec: 459. Anal Calcd. forC₂₈ H₂₉ NO₃ S.1.0HCl: C, 67.80; H, 6.10; N, 2.82. Found: C, 68.06; H, H,6.38; N, 2.60.

Alternate Synthesis of[2-(4-tertbutyloxyphenyl)-3-(4-benzyloxy)phenoxy]benzo[b]thiophenePREPARATION 7 [3-(4-benzyloxy)phenoxy]benzo[b]thiophene-2-boronic acid##STR25##

To a -78° C. solution of [3-(4-benzyloxy)phenoxy]benzo[b]thiophene (5.00g, 15.1 mmol) in 20 mL of anhydrous tetrahydrofuran under N₂ was addedn-butyllithium (9.90 mL, 15.8 mmol, 1.6M in hexanes) dropwise viasyringe. After stirring for 15 minutes, B(OiPr)₃ (3.83 mL, 16.6 mmol)was added via syringe, and the resulting mixture was allowed to warm to0° C. The reaction was then quenched by distributing between ethylacetate and 1.0N hydrochloric acid (100 mL each). The layers wereseparated and the organic was extracted with water (1×100 mL). Theorganic layer was dried (sodium sulfate) and concentrated in vacuo to asolid that was triturated from ethyl ether/hexanes. Filtration provided3.96 g (70%) of [3-(4-benzyloxy)phenoxy]benzo[b]thiophene-2-boronic acidas a white solid. mp 115°-121° C. ¹ H NMR (DMSO-d₆) δ 8.16 (d, J=8.5 Hz,1H), 7.98 (d, J=9.0 Hz, 1H), 7.42-7.23 (m, 7H), 6.90 (q, J_(AB) =9.0 Hz,4H), 5.01 (s, 2H). Anal. Calcd. for C₂₁ H₁₇ O₄ SB: C, 67 04; H, 4.55.Found: C, 67.17; H, 4.78.

[3-(4-Benzyloxy)phenoxy]benzo[b]thiophene-2-boronic acid was reactedwith 4-(-tertbutoxy)bromobenzene according to the conditions describedabove for [2-iodo-3-(4-benzyloxy)phenoxy]benzo[b]thiophene and4-(tertbutoxy)phenyl boronic acid to give[2-(4-tertbutyloxyphenyl)-3-(4-benzyloxy)phenoxy]benzo[b]thiophene in81% yield.

Examples prepared by employing this method are:

EXAMPLE 7[3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(phenyl)]benzo[b]thiophenehydrochloride ##STR26##

mp 223°-226° C. ¹ H NMR (DMSO-d₆) δ 7.99 (d, J=8.2 Hz, 1H), 7.71 (d,J=7.3 Hz, 1H), 7.44-7.30 (m, 7H), 6.90 (s, 4H), 4.27 (m, 2H), 3.43-3.35(m, 4H), 2.97-2.88 (m, 2H), 1.73-1.61 (m, 5H), 1.34 (m, 1H). Anal.Calcd. for C₂₇ H₂₇ NO₂ S.1.0 HCl: C, 69.59; H, 6.06; N, 3.00. Found: C,69.88; H, 6.11; N, 3.19.

EXAMPLE 8[3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-flourophenyl)]benzo[b]thiophene##STR27##

mp 219°-226° C. ¹ H NMR (DMSO-d₆) δ 10.20 (bs, 1H), 7.99 (d, J=8.2 Hz,1H), 7.77-7.73 (m, 4H), 7.42-7.25 (m, 5H), 6.90 (s, 4H), 4.27 (m, 2H),3.44-3.31 (m, 4H), 2.96-2.89 (m, 2H), 1.78-1.61 (m, 5H), 1.34 (m, 1H).FD mass spec: 447. Anal. Calcd. for C₂₇ H₂₆ NO₂ SF.1.0 HCl: C, 67.00; H,5.62; N, 2.89. Found: C, 67.26; H, 5.67; N, 3.03.

PREPARATION 8 Synthesis of[6-Hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophene[6-methoxy-2-(4-methoxyphenyl)-3-bromo]benzo-[b]thiophene ##STR28##

To a solution of [6-methoxy-2-(4-methoxyphenyl)]benzo[b]thiophene (27.0g, 100 mmol) in 1.10 L of chloroform at 60° C. was added bromine (15.98g, 100 mmol) dropwise as a solution in 200 mL of chloroform. After theaddition was complete, the reaction was cooled to room temperature, andthe solvent removed in vacuo to provide 34.2 g (100% ) of[6-methoxy-2-(4-methoxyphenyl)-3-bromo]benzo[b]thiophene as a whitesolid. mp 83°-85° C. ¹ H NMR (DMSO-d₆) δ 7.70-7.62 (m, 4H), 7.17 (dd,J=8.6, 2.0 Hz, 1H), 7.09 (d, J=8.4 Hz, 2H). FD mass spec: 349, 350.Anal. Calcd. for C₁₆ H₁₃ O₂ SBr: C, 55.03; H, 3.75. Found: C, 54.79; H,3.76.

EXAMPLE 9[6-methoxy-2-(4-methoxyphenyl)-3-(4-benzyloxy)phenoxy]-benzo[b]thiophene##STR29##

To a solution of[6-methoxy-2-(4-methoxyphenyl)-3-bromo]benzo[b]thiophene (34.00 g, 97.4mmol) in 60 mL of collidine under N₂ was added 4-benzyloxyphenol (38.96g, 194.8 mmol) and cuprous oxide (14.5 g, 97.4 mmol). The resultantmixture was heated to reflux for 48 hours. Upon cooling to roomtemperature, the mixture was dissolved in acetone (200 mL), and theinorganic solids were removed by filtration. The filtrate wasconcentrated in vacuo, and the residue dissolved in methylene chloride(500 mL). The methylene chloride solution was washed with 3Nhydrochloric acid (3×300 mL), followed by 1N sodium hydroxide (3×300mL). The organic layer was dried (sodium sulfate), and concentrated invacuo. The residue was taken up in 100 mL of ethyl acetate whereupon awhite solid formed that was collected by filtration [recovered[6-methoxy-2-(4-methoxyphenyl)]benzo[b]thiophene (4.62 g, 17.11 mmol].The filtrate was concentrated in vacuo, and then passed through a shortpad of silica gel (methylene chloride as eluant) to remove baselinematerial. The filtrate was concentrated in vacuo, and the residuecrystallized from hexanes/ethyl acetate to provide initially 7.19 g of[6-methoxy-2-(4-methoxyphenyl)-3-(4-benzyloxy)phenoxy]benzo[b]thiopheneas an off-white crystalline solid. The mother liquor was concentratedand chromatographed on silica gel (hexanes/ethyl acetate 80:20) toprovide an additional 1.81 g of product. Total yield of[6-methoxy-2-(4-methoxyphenyl)-3-(4-benzyloxy)phenoxy]benzo[ b]thiophenewas 9.00 g (24% based on recovered starting material). The basic extractwas acidified to pH=4 with 5N hydrochloric acid, and the resultantprecipitate collected by filtration and dried to give 13.3 g ofrecovered 4-benzyloxyphenol. mp 100°-103° C. ¹ H NMR (CDCl₃): δ 7.60 (d,J=8.8 Hz, 2H), 7.39-7.24 (m, 7H), 6.90-6.85 (m, 7H), 4.98 (s, 2H), 3.86(s, 3H) 3.81 (s, 3H). FD mass spec: 468. Anal. Calcd. for C₂₉ H₂₄ O₄ S:C, 74.34; H, 5.16. Found: C, 74.64; H, 5.29.

PREPARATION 9[6-methoxy-2-(4-methoxyphenyl)-3-(4-hydroxy)phenoxy]benzo[b]thiophene##STR30##

To a solution of[6-methoxy-2-(4-methoxyphenyl)-3-(4-benzyloxy)phenoxy]benzo[b]thiophene(1.50 g, 3.20 mmol) in 50 mL of ethyl acetate and 10 mL of 1%concentrate hydrochloric acid in ethanol was added 10%palladium-on-carbon (300 mg). The mixture was hydrogenated at 40 psi for20 minutes, after which time the reaction was judged complete by thinlayer chromatography. The mixture was passed through Celite to removecatalyst, and the filtrate concentrated in vacuo to a white solid. Thecrude product was passed through a pad of silica gel (chloroform aseluant). Concentration provided 1.10 g (91%) of[6-methoxy-2-(4-methoxyphenyl)-3-(4-hydroxy)phenoxy]benzo[b]-thiopheneas a white solid. mp 123°-126° C. ¹ H NMR (DMSO-d₆) d 9.10 (s, 1H), 7.59(d, J=8.8 Hz, 2H), 7.52 (d, J=2.1 Hz, 1H), 7.14 (d, J=8.8 Hz, 1H), 6.95(d, J=8.8 Hz, 2H), 6.89 (dd, J=8.8, 2.1 Hz, 1H), 6.72 (d, J=9.0 Hz, 2H),6.63 (d, J=9.0 Hz, 2H), 3.78 (s, 3H), 3.72 (s, 3H). FD mass spec: 378.Anal. Calcd. for C₂₂ H₁₈ O₄ S: C, 69.82; H, 4.79. Found: C, 70.06; H,4.98.

EXAMPLE 10[6-methoxy-3-[4-[2-(1-piperidinyl)ethoxy]-phenoxy]-2-(4-methoxyphenyl)]benzo[b]thiophene.##STR31##

To a solution of[6-methoxy-2-(4-methoxyphenyl)-3-(4-hydroxy)phenoxy]benzo[b]thiophene(1.12 g, 2.97 mmol) in 7 mL of anhydrous N,N-dimethylformamide under N₂was added cesium carbonate (3.86 g, 11.88 mmol). After stirring for 10minutes, 2-chloroethylpiperidine hydrochloride (1.10 g, 1.48 mmol) wasadded. The resultant mixture was stirred for 18 hours at ambienttemperature. The reaction was the distributed between chloroform/water(100 mL each). The layers were separated and the aqueous extracted withchloroform (3×50 mL). The organic was combined and washed with water(2×100 mL). Drying of the organic (sodium sulfate) and concentrationprovided an oil that was chromatographed on silica gel (2%methanol/chloroform). The desired fractions were concentrated to an oilthat was dissolved in 10 mL of ethyl acetate and treated with oxalicacid (311 mg, 3.4 mmol). After stirring for 10 minutes, a whiteprecipitate formed and was collected by filtration and dried to provide1.17 g (70%) overall of[6-methoxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-methoxyphenyl)]benzo[b]thiopheneas the oxalate salt. mp 197°-200° C. (dec). ¹ H NMR (DMSO-d₆) δ 7.60 (d,J=8.7 Hz, 2H), 7.55 (d, J=1.1 Hz, 1H), 7.14 (d, J=8.8 Hz, 1H), 7.06 (d,J=8.8 Hz, 2H), 6.91 (dd, J=8.8, 1.1 Hz, 1H), 6.87 (s, 4H), 4.19 (broadt, 2H), 3.78 (s, 3H), 3.72 (s, 3H), 3.32 (broad t, 2H), 3.12-3.06 (m,4H), 1.69-1.47 (m, 4H), 1.44-1.38 (m, 2H). FD mass spec: 489. Anal.Calcd. for C₂₉ H₃₁ NO₄ S.0.88 HO₂ CCO₂ H: C, 64.95; H, 5.80; N, 2.46.Found: C, 64.92; H, 5.77; N, 2.54.

EXAMPLE 11 Treatment of Free Base with Ethyl Ether.hydrochloric AcidProvided[6-methoxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-methoxyphenyl)]benzo[b]thiophenehydrochloride ##STR32##

mp 216°-220° C. ¹ H NMR (DMSO-d₆) δ 10.20 (bs, 1H), 7.64 (d, J=8.7 Hz,2H), 7.59 (d, J=1.5 Hz, 1H), 7.18 (d, J=9.0 Hz, 1H), 7.00 (d, J=8.7 Hz,1H), 6.96 (dd, J=9.0, 1.5 Hz, 1H), 6.92 (q, J_(AB) =9.0 Hz, 4H), 4.31(m, 2H), 3.83 (s, 3H), 3.77 (s, 3H), 3.43 (m, 4H), 2.97 (m, 2H), 1.77(m, 5H), 1.37 (m, 1H). FD mass spec: 489. Anal. Calcd. for C₂₉ H₃₁ NO₄S.1.0 HCl: C, 66.21; H, 6.13; N, 2.66. Found: C, 66.,46; H, 6.16; N,2.74.

Prepared in an analogous manner were the following examples:

EXAMPLE 12[6-Methoxy-3-[4-[2-(1-pyrolodinyl)ethoxy]phenoxy]-2-(4-methoxyphenyl)]benzo[b]thiophene##STR33##

mp 95°-98° C. ¹ H NMR (DMSO-d₆) δ 7.64 (d, J=9.0 Hz, 2H), 7.58 (d, J=2.0Hz, 1H), 7.18 (d, J=9.0 Hz, 1H), 7.00 (d, J=9.0 Hz, 2H), 6.94 (dd,J=9.0, 2.0 Hz, 1H), 6.86 (s, 4H), 3.97 (t, J=6.0 Hz, 2H), 3.83 (s, 3H),3.76 (s, 3H), 2.73 (t, J=6.0 Hz, 2H), 2.51 (m, 4H), 1.66 (m, 4H). FDmass spec: 477. Anal. Calcd. for C₂₈ H₂₉ NO₄ S: C, 70.71; H, 6.15; N,2.99. Found: C, 70.59; H, 6.15; N, 3.01.

EXAMPLE 13 [6-Methoxy-3-[4-[2-(1-hexamethyleneimino)ethoxy]phenoxy]-2-(4-methoxyphenyl)]benzo[b]thiophene hydrochloride ##STR34##

mp 189°-192° C. ¹ H NMR (DMSO-d₆) δ 10.55 (bs, 1H), 7.64 (d, J=9.0 Hz,2H), 7.58 (d, J=2.0 Hz, 1H), 7.19 (d, J=9.0 Hz, 1H), 7.00 (d, J=9.0 Hz,2H), 6.95 (dd, J=9.0, 2.0 Hz, H), 6.86 (s, 4H), 3.94 (t, J=6.0 Hz, 2H),3.83 (s, 3H), 3.76 (s, 3H), 2.80 (t, J=6.0 Hz, 2H), 2.66 (m, 4H), 1.53(m, 8H). Anal. Calcd. for C₃₀ H₃₃ NO₄ S.1.0 HCl: C, 66.71; H, 6.35; N,2.59. Found: C, 66.43; H, 6.46; N, 2.84.

EXAMPLE 14[6-Methoxy-3-[4-[2-(1-N,N-diethylamino)ethoxy]phenoxy]-2-(4-methoxyphenyl)]benzo[b]thiophenehydrochloride ##STR35##

mp 196°-198° C. ¹ H NMR (DMSO-d₆) δ 10.48 (bs, 1H), 7.64 (d, J=9.0 Hz,2H), 7.59 (d, J=2.0 Hz, 1H), 7.19 (d, J=9.0 Hz, 1H), 7.00 (d, J=9.0 Hz,2H), 6.97 (dd, J=9.0, 2.0 Hz, 1H), 6.87 (q, J_(AB) =9.0 Hz, 4H), 4.25(m, 2H), 3.83 (s, 3H), 3.77 (s, 3H), 3.54 (m, 2H), 3.09 (m, 4H), 2.00(m, 3H), 1.88 (m, 3H). Anal. Calcd. for C₂₈ H₃₁ NO₄ S.1.5 HCl: C, 63.18;H, 6.15; N, 2.63. Found: C, 63.46; H, 5.79; N, 2.85.

EXAMPLE 15 [6-Methoxy-3-[4-[2-(morpholino)ethoxy]phenoxy]-2-(4-methoxyphenyl)]benzo[b]thiophenehydrochloride ##STR36##

mp 208°-211° C. ¹ H NMR (DMSO-d₆) δ 10.6 (bs, 1H), 7.63 (d, J=9.0 Hz,2H), 7.60 (d, J=2.0 Hz, 1H), 7.20 (J=9.0 Hz, 1H), 7.00 (d, J=9.0 Hz,2H), 6.97 (dd, J=9.0, 2.0 Hz, 1H), 6.91 (q, J_(AB) =9.0 Hz, 4H), 4.29(m, 2H), 4.08-3.91 (m, 4H), 3.82 (s, 3H), 3.77 (s, 3H), 3.59-3.42 (m,4H), 3.21-3.10 (m, 2H). Anal. Calcd. for C₂₈ H₂₉ NO₅ S.1.0 HCl: C,63.09; H, 5.73; N, 2.65. Found: C, 63.39; H, 5.80; N, 2.40.

EXAMPLE 16[6-Methoxy-3-[4-[3-(piperidino)propoxy]phenoxy]-2-(4-methoxyphenyl)]benzo[b]thiophenehydrochloride ##STR37##

mp 195°-200° C. ¹ H NMR (DMSO-d₆) δ 9.90 (bs, 1H), 7.64 (d, J=9.0 Hz,2H), 7.59 (d, J=2.0 Hz, 1H), 7.18 (d, J=9.0 Hz, 1H), 7.00 (d, J=9.0 Hz,2H), 6.95 (dd, J=9.0, 2.0 Hz, 1H), 6.88 (s, 4H), 3.97 (t, J=6.0 Hz, 2H),3.83 (s, 3H), 3.77 (s, 3H), 3.44 (m, 2H), 3.15 (m, 2H), 2.87 (m, 2H),2.12 (m, 2H), 1.77 (m, 5H), 1.39 (m, 1H). Anal. Calcd. for C₃₀ H₃₃ NO₄S.1.15 HCl: C, 66.01; H, 6.40; N, 2.73. Found: C, 66.01; H, 6.40; N,2.73.

EXAMPLE 17[6-Methoxy-3-[4-[3-(1-N,N-diethylamino)propoxy]phenoxy]-2-(4-methoxyphenyl)]benzo[b]thiophenehydrochloride ##STR38##

mp 164°-166° C. ¹ H NMR (DMSO-d₆) δ 9.77 (bs, 1H), 7.64 (d, J=9.0 Hz,2H, 7.59 (d, J=2.0 Hz, 1H), 7.18 (d, J=9.0 Hz, 1H), 7.00 (d, J=9.0 Hz,2H), 6.95 (dd, J=9.0, 2.0 Hz, 1H), 6.89 (s, 4H), 3.99 (t, J=6.0 Hz,2H),3.83 (s, 3H), 3.77 (s, 3H), 3.15 (m, 6H), 2.06 m, 2H), 1.20 (t, J=7.0Hz, 6H). Anal. Calcd. for C₂₉ H₃₃ NO₄ S.1.0 HCl: C, 65.96; H, 6.49; N,2.65. Found: C, 66.25; H, 6.64; N, 2.84.

EXAMPLE 18 [6-hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]-phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophene##STR39##

[6-methoxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-methoxyphenyl)]benzo[b]thiophenehydrochloride (10.00 g, 19.05 mmol) was dissolved in 500 mL of anhydrousmethylene chloride and cooled to 8° C. To this solution was added borontribromide (7.20 mL, 76.20 mmol). The resultant mixture was stirred at8° C. for 2.5 hours. The reaction was quenched by pouring into astirring solution of saturated sodium bicarbonate (1 L), cooled to 0° C.The methylene chloride layer was separated, and the remaining solidswere dissolved in methanol/ethyl acetate. The aqueous layer was thenextracted with 5% methanol/ethyl acetate (3×500 mL). All of the organicextracts (ethyl acetate and methylene chloride) were combined and dried(sodium sulfate). Concentration in vacuo provided a tan solid that waschromatographed (silicon dioxide, 1-7% methanol/chloroform) to provide7.13 g (81%) of[6-hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]-thiopheneas a white solid. mp 93° C. ¹ H NMR (DMSO-d₆) δ 9.73 (bs, 1H), 9.68 (bs,1H), 7.45 (d, J= 8.6 Hz, 2H), 7.21 (d, J=1.8 Hz, 1H), 7.04 (d, J=8.6 Hz,1H), 6.84 (dd, J=8.6, 1.8 Hz, 1H (masked)), 6.81 (s, 4H), 6.75 (d, J=8.6Hz, 2H), 3.92 (t, J=5.8 Hz, 2H), 2.56 (t, J=5.8 Hz, 2H), 2.36 (m. 4H),1.43 (m, 4H), 1.32 (m, 2H). FD mass spec: 462. Anal. Calcd. for C₂₇ H₂₇NO₄ S: C, 70.20; H, 5.90; N, 3.03. Found: C, 69.96; H, 5.90; N, 3.14.

EXAMPLE 19[6-hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiopheneis Converted to its Oxalate Salt in 80% Yield by the Procedure DescribedAbove Data for [6-hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiopheneoxalate ##STR40##

mp 246°-249° C. (dec). ¹ H NMR (DMSO-d₆) δ 7.45 (d, J=8.6 Hz, 2H), 7.22(d, J=1.8 Hz, 1H), 7.05 (d, J=8.6 Hz, 1H), 6.87 (dd, J=8.6, 1.8 Hz, 1H(masked)), 6.84 (s, 4H), 6.75 (d, J=8.6 Hz, 2H), 4.08 (bt, 2H), 3.01(bt, 2H), 2.79 (m, 4H), 1.56 (m, 4H), 1.40 (m, 2H). FD mass spec 462.Anal. Calcd. for C₂₇ H₂₇ NO₄ S.0.75 HO₂ CCO₂ H: C, 64.63; H, 5.42; N,2.64. Found: C, 64.61; H, 5.55; N, 2.62.

EXAMPLE 20[6-hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophenewas Converted to its Hydrochloride Salt in 91% Yield by Treatment of theFree Base in Ethyl Acetate with Ethyl Ether.hydrochloric Acid Data for[6-hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophenehydrochloride ##STR41##

mp 158°-165° C. ¹ H NMR (DMSO-d₆) δ 9.79 (s, 1H), 9.74 (s, 1H), 7.40 (d,J=8.6 Hz, 2H), 7.23 (d, J=2.0 Hz, 1H), 7.04 (d, J=8.6 Hz, 1H), 6.86 (q,J_(AB) =9.3 Hz, 4H), 6.76 (dd, J=8.6, 2.0 Hz, 1), 6.74 (d, J=8.6 Hz,2H), 4.26 (bt, 2H), 3.37 (m, 4H), 2.91 (m, 2H), 1.72 (m, 5 H), 1.25 (m,1H). FD mass spec 461. Anal. Calcd. for C₂₇ H₂₇ NO₄ S.1.0 HCl: C, 65.11;H, 5.67; N, 2.81. Found: C, 64.84; H, 5.64; N, 2.91.

Prepared in an analogous manner were the following examples:

EXAMPLE 21[6-hydroxy-3-[4-[2-(1-pyrolidinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophene##STR42##

mp 99°-113° C. ¹ H NMR (DMSO-d₆) δ 9.75 (s, 1H), 9.71 (s, 1H), 7.50 (d,J=9.0 Hz, 2H), 7.25 (d, J=2.0 Hz, 1H), 7.09 (d, J=9.0 Hz, 1H), 6.85 (s,1H), 6.80 (dd, J=9.0, 2.0 Hz, 1H), 6.79 (d, J=9.0 Hz, 2H), 3.93 (m, 2H),2.73 (m, 2H), 2.53 (m, 4H), 0.96 (t, J=7.0 Hz, 4H). Anal. Calcd. for C₂₆H₂₅ NO₄ S.0.5 H₂ O: C, 68.40; H, 5.74; N, 3.07. Found: C, 68.52; H,6.00; N, 3.34.

EXAMPLE 22[6-hydroxy-3-[4-[2-(1-hexamethyleneimino)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophene ##STR43##

mp 125°-130° C. ¹ H NMR (DMSO-d₆) δ 9.75 (s, 1H), 9.71 (s, 1H), 7.50 (d,J=9.0 Hz, 2H), 7.26 (d, J=2.0 Hz, 1H), 7.09 (d, J=9.0 Hz, 1H), 6.85 (s,3H), 6.80 (dd, J=9.0, 2.0 Hz, 1H), 6.79 (d, J=9.0 Hz), 3.94 (t, J=6.0Hz, 2H), 2.80 (t, J=6.0 Hz, 2H), 2.66 (m, 4H), 1.53 (m, 8H). Anal.Calcd. for C₂₈ H₂₉ NO₄ S: C, 70.71; H, 6.15; N, 2.94. Found: C, 70.67;H, 6.31; N, 2.93.

EXAMPLE 23[6-hydroxy-3-[4-[2-(1-N,N-diethylamino)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophene##STR44##

mp 137°-141° C. ¹ H NMR (DMSO-d₆) δ 9.75 (s, 1H), 9.71 (s, 1H), 7.49 (d,J=9.0 Hz, 1H), 7.25 (d, j=2.0 Hz, 1H), 7.09 (d, J=9.0 Hz, 1H), 6.85 (s,4H), 6.80 (dd, J=9.0, 2.0 Hz, 1H), 6.79 (d, J=9.0 Hz, 2H), 3.95 (t,J=6.0 Hz, 2H), 2.74 (t, J=6.0 Hz, 2H), 2.51 (m, 4H), 1.66 (m, 6H). Anal.Calcd. for C₂₆ H₂₇ NO₄ S: C, 69.46; H, 6.05; N, 3.12. Found: C, 69.76;H, 5.85; N, 3.40.

EXAMPLE 24[6-hydroxy-3-[4-[2-(morpholino)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophenehydrochloride ##STR45##

mp 157°-162° C. ¹ H NMR (DMSO-d₆) δ 10.60 (bs, 1H), 9.80 (s, 1H), 9.75(s, 1H), 7.50 (d, J=9.0 Hz, 2H), 7.28 (d, J=2.0 Hz, 1H), 7.10 (d, J=9.0Hz, 1H), 6.92 (q, J_(AB) =9.0 Hz, 4H), 6.81 (dd, J=9.0, 2.0 Hz, 1H),6.80 (d, J=9.0 Hz, 2H), 4.30 (m, 2H), 3.95 (m, 2H), 3.75 (m, 2H), 3.51(m, 4H), 3.18 (m, 2H). Anal. Calcd. for C₂₆ H₂₅ NO₅ S.HCl: C, 62.46; H,5.24; N, 2.80. Found: C, 69.69; H, 5.43; N, 2.92.

EXAMPLE 25[6-hydroxy-3-[4-[3-(1-N,N-diethylamino)propoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophenehydrochloride ##STR46##

mp 185°-191° C. ¹ H NMR (DMSO-d₆) δ 9.94 (bs, 1H), 9.81 (s, 1H), 9.75(s, 1H), 7.50 (d, J=9.0 Hz, 2H), 7.27 (dd, J=2.0 Hz, 1H), 7.10 (d, J=9.0Hz, 1H), 6.87 (s, 4H), 6.80 (dd, J=9.0, 2.0 Hz, 1H), 6.79 (d, J=9.0 Hz,2H), 3.99 (t, J=6.0 Hz, 2H), 3.14 (m, 6H), 2.08 (m, 2H), 1.20 (t, J=6.0Hz, 6H). Anal. Calcd. for C₂₇ H₂₉ NO₄ S.1.30HCl: C, 63.46; H, 5.98; N,2.74. Found: C, 63.23; H, 6.03; N, 3.14.

EXAMPLE 26 [6-hydroxy-3-[4-[2-(1-N,N-diisopropylamino)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophenehydrochloride ##STR47##

mp 128°-131° C. ¹ H NMR (DMSO-d₆) δ 9.81 (bs, 1H), 9.76 (s, 1H), 9.02(s, 1H), 7.49 (d, J=9.0 Hz, 2H), 7.28 (m, 1H), 7.09 (d, J=9.0 Hz, 1H),6.90 (s, 4H), 6.79 (m, 3H), 4.19 (m, 2H), 3.68 (m, 2H), 3.50 (m, 2H).1.31 (m, 12H). Anal. Calcd. for C₂₈ H₃₁ NO₄ S.1.33HCl: C, 63.92; H,6.19; N, 2.66. Found: C, 63.82; H, 6.53; N, 2.61.

EXAMPLE 27[6-hydroxy-3-[4-[3-(piperidino)propoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophenehydrochloride ##STR48##

mp 258°-262° C. ¹ H NMR (DMSO-d₆) δ 9.85 (bs, 1H), 9.81 (s, 1H), 9.75(s, 1H), 7.50 (d, J=9.0 Hz, 2H), 7.27 (d, J=2.0 Hz, 1H), 7.10 (d, J=9.0Hz, 1H), 6.87 (s, 4H), 6.80 (dd, J=9.0, 2.0 Hz, 1H), 6.79 (d, J=9.0 Hz,2H), 3.97 (t, J=6.0 Hz, 2H), 3.44 (m, 2H), 3.15 (m, 2H), 2.88 (m, 2H),2.11 (m, 2H), 1.73 (m, 5H), 1.39 (m, 1H). Anal. Calcd. for C₂₈ H₂₉ NO₄S.0.75HCl: C, 66.87; H, 5.96; N, 2.78. Found: C, 67.04; H, 5.90; N,2.68.

Alternatively, as shown in Scheme III, supra, Example 19 was preparedusing the methoxymethyl (MOM) protecting groups in place of methoxy. Themethods are directly analogous to those just described, with theexception that the MOM groups are removed in the final step by acidhydrolysis.

PREPARATION 10[6-methoxy-2-(4-methoxmethyloxyphenyl)-3-(4-benzyloxy)phenoxy]benzo[b]thiophene##STR49##

mp 94°-96° C. ¹ H NMR (DMSO-d₆) δ 7.65 (d, J=2.0 Hz, 1H), 7.64 (d, J=8.6Hz, 2H), 7.43-7.32 (m, 5H), 7.23 (d, J=8.8 Hz, 1H), 7.08 (d, J=8.6 Hz,2H), 7.04 (dd, J=8.8, 2.0 Hz, 1H), 6.92 (q, J_(AB) =9.2 Hz, 4H), 5.26(s, 2H), 5.21 (s, 2H), 5.01 (s, 3H), 3.40 (s, 3H), 3.37 (s, 3H). FD massspec 528.

PREPARATION 11[6-methoxy-2-(4-methoxmethyloxyphenyl)-3-(4-hydroxy)phenoxy]benzo[b]thiophene##STR50##

mp 90°-91° C. ¹ H NMR (DMSO-d₆) δ 9.15 (s, 1H), 7.65 (d, J=8.1 Hz, 2H),7.63 (d, J=2.0 Hz, 1H), 7.22 (d, J=8.8 Hz, 1H), 7.05 (dd, J=8.8, 2.0 Hz,1H), 6.72 (q, J_(AB) =9.1 Hz, 4H), 5.26 (s, 2H), 5.21 (s, 2H), 3.40 (s,3H), 3.37 (s, 3H). FD mass spec 438. Anal. Calcd. for C₂₄ H₂₂ O₆ S: C,65.74; H, 5.06. Found: C, 65.50; H, 4.99.

EXAMPLE 29[6-benzoyloxy-3-[4-[2-(1-piperidinyl)-ethoxy]phenoxy]-2-(4-benzoyloxyphenyl)]benzo[b]thiophenehydrochloride ##STR51##

To a solution of Example 20 (0.50 g, 1.08 mmol) in 20 mL of anhydroustetrahydrofuran at 0° C. was added triethylamine (1.00 mL). To thismixture was added benzoylchloride (0.28 mL, 2.35 mmol). After stirringat 0° C. for 2 hours, the reaction was quenched by distributing betweenethyl acetate/saturated sodium bicarbonate solution (100 mL each). Thelayers were separated and the organic was dried (sodium sulfate) andconcentrated in vacuo to a white solid. The crude product was dissolvedin 10 mL of ethyl acetate and treated with ethyl ether.hydrochloricacid. A white precipitate formed that was collected by filtration.Drying provided 390 mg (50%) of [6-benzoyloxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-benzoyloxyphenyl)]benzo[b]thiophene hydrochlorideas a white solid. mp 200°-204° C. ¹ H NMR (DMSO-d₆) δ 9.95 (bs, 1H),8.18 (m, 1H), 8.16 (m, 2H), 8.12 (dd, J=10.0, 2.0 Hz, 2H), 7.87 (dd,J=7.0, 2.0 Hz, 2H), 7.78 (m, 2H), 7.64 (m, 2H), 7.42 (d, J=7.0 Hz, 2H),7.34 (dd, J=8.0, 2.0 Hz, 1H), 7.00 (s, 4H), 4.32 (m, 2H), 3.45 (m, 4H),2.99 (m, 2H), 1.75 (m, 5H), 1.39 (m, 1H). Anal. Calcd. for C₄₁ H₃₅ NO₆S.1.5HCl: C, 67.97; H, 5.08; N, 1.93. Found: C, 68.05; H, 5.24; N, 2.01.

By the same procedure was prepared:

EXAMPLE 30[6-ethylsulfonyloxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-ethylsulfonyloxyphenyl)]benzo[b]thiophenehydrochloride ##STR52##

Yield=72%. mp 110°-115° C. ¹ H NMR (DMSO-d₆) δ 10.15 (bs, 1H), 8.15 (d,J=2.0 Hz, 1H), 7.85 (d, J=7.0 Hz, 2H), 7.43 (m, 3H), 7.34 (dd, J=9.0,2.0 Hz, 1H), 6.97 (m, 4H), 4.31 (m, 2H), 3.57 (m, 4H), 3.44 (m, 4H),2.97 (m, 2H), 1.76 (m, 5H), 1.40 (m, 7H). Anal. Calcd. for C₃₁ H₃₅ NO₈S₃.1.5HCl: C, 54.57; H, 5.32; N, 2.05. Found: C, 54.36; H, 5.37; N,2.05.

Prepared from Example 1 by similar procedures were:

EXAMPLE 313-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-benzoyloxyphenyl)]benzo[b]thiophenehydrochloride ##STR53##

Yield=85%. mp 190°-198° C. ¹ H NMR (DMSO-d₆) δ 10.48 (br s, 1H),8.00-8.10 (m, 2H), 7.80-8.00 (m, 3H), 7.60-7.53 (m, 4H), 7.40-7.56 (m,6H), 6.93 (s, 2H), 4.37-4.43 (m, 2H), 3.00-3.05 (m, 2H), 2.53-2.63 (m,6H), 1.75-1.95 (m, 3H), 1.40-1.50 (m, 1H). FD mass spec: 550. Anal.Calcd. for C₃₄ H₃₁ NO₄ S.1.0HCl: C, 74.29; H, 5.68; N, 2.55. Found: C,74.52; H, 5.80; N, 2.59.

EXAMPLE 323-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-pivaloyloxyphenyl)]benzo[b]thiophenehydrochloride ##STR54##

Yield=90%. mp=193°-197° C. ¹ H NMR (DMSO-d₆) δ 10.10 (br s, 1H), 8.12(d, J=8.0 Hz, 1H), 7.85 (d, J=8.6 Hz, 1H), 7.40-7.53 (m, 3H), 7.15 (d,J=6.7 Hz, 2 H), 7.00 (s, 5H), 4.33-4.40 (m, 2H), 3.45-3.60 (m, 4H),3.00-3.10 (m, 2H), 1.70-1.90 (m, 6H), 1.40 (s, 9H). FD mass spec: 529.Anal. Calcd. for C₃₂ H₃₅ NO₄ S.1.0HCl: C, 67.89; H, 6.41; N, 2.47.Found: C, 68.94; H, 6.61; N, 1.72.

EXAMPLE 333-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-butylsulfonyloxyphenyl)]benzo[b]thiophenehydrochloride ##STR55##

Yield=85% white solid. mp=98°-104° C. ¹ H NMR (DMSO-d₆) δ 10.20 (br s,1H), 8.02 (d, J=8.0 Hz, 1H), 7.82 (d, J=8.7 Hz, 2H), 7.40-7.55 (m, 5H),7.00 (s, 4H), 4.30-4.40 (m, 2H), 3.46-3.66 (m, 6H), 3.00-3.10 (m, 2H),1.70-1.95 (m, 6H), 1.40-1.60 (m, 4H), 0.87 (t, J=7.3 Hz, 3H). FD massspec: 565. Anal. Calcd. for C₃₁ H₃₅ NO₅ S₂.1.0HCl: C, 61.83; H, 6.03; N,2.33. Found: C, 61.55; H, 6.15; N, 2.25.

PREPARATION 12[6-hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]-thiophenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophene##STR56##

Preparation of 4-(methoxymethyloxy)phenyldisulfide.

To a solution of 4-hydroxyphenyldisulfide (650 mg, 2.60 mmol) in 10 mLof anhydrous N,N-dimethylformamide at 10° C. was added sodium hydride(230 mg, 5.75 mmol, 60% dispersion in mineral oil). After stirring for15 minutes, chloromethylmethyl ether (0.44 mL, 5.75 mmol) was added viasyringe. The reaction was warmed to ambient temperature and stirred for0.5 hours. The mixture was distributed between brine/ethyl acetate (20mL each). The layers were separated and the aqueous phase extracted withethyl acetate (2×20 mL). The organic was dried (sodium sulfate) andconcentrated to a yellow oil (993 mg, 100%). An analytical sample of4-(methoxymethyloxy)-phenyldisulfide was prepared by chromatography(silicon dioxide, 4% ethyl acetate/hexanes). ¹ H NMR (DMSO-d₆) δ 7.40(d, J=6.9 Hz, 4H), 7.00 (d, J=6.9 Hz, 4H), 5.15 (s, 4H), 3.32 (s, 6H).FD mass spec: 338. Anal. Calcd. for C₁₆ H₁₈ O₄ S₂ : C, 56.78; H, 5.36.Found: C, 57.08; H, 5.44.

PREPARATION 13[6-methoxy-2-(4-methoxyphenyl)-3-(4-methoxymethyleneoxy)thiophenoxy]benzo[b]thiophene##STR57##

To a solution of[6-methoxy-2-(4-methoxyphenyl)-3-bromo]benzo[b]thiophene (1.82 g, 5.2mmol) in 10 mL of anhydrous tetrahydrofuran under N₂ at -60° C. wasadded n-butyllithium (3.15 mL, 5.0 mmol, 1.6M solution in hexanes)dropwise via syringe. The resulting mixture was warmed to -20° C. for 10minutes, then cooled back to -60° C.4-(methoxymethyloxy)-phenyldisulfide (800 mg, 2.36 mmol) in 5 mL ofanhydrous tetrahydrofuran was added to the lithio species, and theresultant mixture was allowed to gradually warm to 0° C. After stirringfor 20 minutes, the reaction was quenched by distributing betweenbrine/ethyl acetate (50 mL each). The layers were separated, and theaqueous phase was extracted with ethyl acetate (2×50 mL). The organiclayer was combined, dried (sodium sulfate), and concentrated in vacuo toan oil. Chromatography (silicon dioxide, 5% ethyl acetate/hexanes)provided 287 mg (27%) of[6-methoxy-2-(4-methoxyphenyl)-3-(4-methoxymethyleneoxy)thiophenoxy]benzo[b]thiopheneas a colorless oil. ¹ H NMR (DMSO-d₆) δ 7.59 (d, J=8.4 Hz, 2H), 7.58 (d,J=2.0 Hz, 1H), 7.52 (d, J=8.8 Hz, 1H), 7.03-6.85 (m, 7H), 5.06 (s, 2H),3.79 (s, 3H), 3.76 (s, 3H). FD mass spec: 438. Anal. Calcd. for C₂₄ H₂₂O₄ S₂ : C, 65.73; H, 5.06. Found: C, 65.93; H, 5.10.

PREPARATION 14[6-methoxy-2-(4-methoxyphenyl)-3-(4-hydroxy)thiophenoxy]benzo[b]thiophene##STR58##

To a solution of [6-methoxy-2-(4-methoxyphenyl)-3-(4-methoxymethyleneoxy)thiophenoxy]benzo[b]thiophene (233 mg, 0.53mmol) in 10 mL of a 1:1:2 mixture of methanol:water:tetrahydrofuran wasadded methane sulfonic acid (0.2 mL, 2.66 mmol). The mixture was heatedto reflux for 5 hour. Upon cooling to ambient temperature, the reactionmixture was diluted with water. The aqueous phase was extracted withethyl acetate (2x). The organic layer was washed with sat sodiumbicarbonate solution several times. The organic layer was dried (sodiumsulfate) and concentrated in vacuo to provide 206 mg (99%) of[6-methoxy-2-(4-methoxyphenyl)-3-(4-hydroxy)thiophenoxy]benzo-[b]thiopheneas a colorless oil. ¹ H NMR (DMSO-d₆) δ 9.43 (s, 1H), 7.63 (d, J=8.4 Hz,2H), 7.61 (d, J=2.0 Hz, 1H), 7.59 (d, J=8.8 Hz, 1H), 7.08 (d, J= 8.4 Hz,2H), 7.02 (dd, J=8.8, 2.0 Hz, 1H), 6.90 (d, J=8.6 Hz, 2H), 6.63 (d,J=8.6 Hz, 2H). FD mass spec: 395. Anal. Calcd. for C₂₂ H₁₈ O₃ S₂ : C,66.98; H, 4.60. Found: C, 67.26; H, 4.78.

EXAMPLE 34[6-methoxy-3-[4-[2-(1-piperidinyl)ethoxy]thiophenoxy]-2-(4-methoxyphenyl)]benzo[b]thiophene##STR59##

To a solution of[6-methoxy-2-(4-methoxyphenyl)-3-(4-hydroxy)thiophenoxy]benzo[b]thiophene(242 mg, 0.61 mmol) in 8.0 mL of anhydrous N,N-dimethylformamide wasadded cesium carbonate (820 mg, 2.5 mmol) followed by2-chloroethylpiperidine hydrochloride (194 mg, 1.05 mmol). The resultingmixture was stirred for 48 hours at ambient temperature and thendistributed between brine/ethyl acetate. The layers were separated, andthe aqueous phase was extracted with ethyl acetate (3x). The organiclayer was dried (sodium sulfate) and concentrated in vacuo to an oil.Chromatography (silicon dioxide, 0-2% methanol/chloroform) provided 244mg (92%) of[6-methoxy-3-[4-[2-(1-piperidinyl)ethoxy]thiophenoxy]-2-(4-methoxyphenyl)]benzo[b]thiopheneas an amber oil.

EXAMPLE 35 A Sample of[6-methoxy-3-[4-[2-(1-piperidinyl)ethoxy]thiophenoxy]-2-(4-methoxyphenyl)]benzo[b]thiophenewas Converted to its Hydrochloride Salt According to the StandardProcedure in 72% Yield ##STR60##

mp 198°-201° C. ¹ H NMR (DMSO-d₆) δ 7.63 (d, J=8.6 Hz, 2H), 7.62 (d,J=2.0 Hz, 1H), 7.58 (d, J=8.2 Hz, 1H), 7.07 (d, J=8.6 Hz, 2H), 7.02 (dd,J=8.2, 2.0 Hz, 1H), 6.92 (q, J_(AB) =9.0 Hz, 4H), 4.24 (bt, 2H), 3.82(s, 3H), 3.80 (s, 3H), 3.49-3.39 (m, 4H), 2.93 (m, 2H), 1.82-1.62 (m,5H), 1.38 (m, 1H). Anal. Calcd. for C₂₉ H₃₂ NO₃ S₂.1.0 HCl: C, 64.28; H,5.95; N, 2.58. Found: C, 64.09; H, 6.08; N, 2.78.

EXAMPLE 36 [6-hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]-thiophenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophene##STR61##

To a solution of[6-methoxy-3-[4-[2-(1-piperidinyl)ethoxy]thiophenoxy]-2-(4-methoxyphenyl)]benzo[b]thiophenehydrochloride (160 mg, 0.29 mmol) in 15 mL of anhydrous methylenechloride at 0° C. under N₂ was added boron tribromide (0.15 mL). Theresulting dark solution was stirred for 1 hour at 0° C. and thenimmediately poured into a stirred solution of ethyl acetate/sat sodiumbicarbonate solution (50 mL each). The layers were separated, and theaqueous phase was washed with ethyl acetate (3×30 mL). The organic wasdried (sodium sulfate) and concentrated in vacuo to a white solid.Chromatography (silicon dioxide, 0-5% methanol/chloroform) provided 91mg (60%) of[6-hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]thiophenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiopheneas a white solid. mp 123°-127° C. ¹ H NMR (DMSO-d₆) δ 9.79 (s, 1H), 9.71(s, 1H), 7.46 (d, J=8.4 Hz, 2H), 7.42 (d, J=8.9 Hz, 1H), 7.26 (d, J=2.0Hz, 1H), 6.91 (d, J=8.8 Hz,2H), 6.82-6.76 (m, 5H), 3.91 (t, J=8.8 Hz,2H), 2.56 (t, J=5.8 Hz, 2H), 2.40 (m, 4H), 1.41-1.28 (m, 6H). FD massspec: 478. Anal. Calcd. for C₂₇ H₂₇ NO₃ S₂ : C, 67.90; H, 5.70; N, 2.93.Found: C, 68.14; H, 5.84; N, 2.65.

EXAMPLE 37[6-Hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]thiophenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophenehydrochloride ##STR62##

mp 180°-190° C. ¹ H NMR (DMSO-d₆) δ 9.86 (s, 1H), 9.79 (s, 1H), 7.46 (d,J=8.5 Hz, 2H), 7.42 (d, J=8.7 Hz, 1H), 7.29 (d, J=2.0 Hz, 1H), 6.96 (d,J=8.7 Hz, 2H), 6.86-6.81 (m, 5H), 4.27 (m, 2H), 3.41-3.37 (m, 4H),2.96-2.84 (m, 2H), 1.77-1.60 (m, 5H), 1.35-1.28 (m, 1H). FD mass spec:477. Anal. Calcd. for C₂₇ H₂₇ NO₃ S₂.2.2 HCl: C, 58.13; H, 5.28; N,2.51. Found: C, 58.11; H, 5.10; N, 2.61.

Prepared by the same procedures were:

EXAMPLE 38[6-methoxy-3-[4-[2-(1-pyrolodinyl)ethoxy]thiophenoxy]-2-(4-methoxyphenyl)]benzo[b]thiophenehydrochloride ##STR63##

mp 215°-218° C. ¹ H NMR (DMSO-d₆) δ 7.61-7.58 (m, 3H), 7.52 (d, J=8.8Hz, 1H), 7.04-6.95 (m, 5H), 6.86 (d, J=8.8 Hz, 2H), 4.22 (bt, 2H), 3.79(s, 3H), 3.76 (s, 3H), 3.47-3.42 (m, 4H), 3.01 (m, 2H), 1.94-1.80 (m,4H). FD mass spec: 491. Anal. Calcd. for C₂₈ H₂₉ NO₃ S₂.1.0HCl: C,63.67; H, 5.73; N, 2.65. Found: C, 63.47; H, 5.78; N, 2.65.

EXAMPLE 39[6-hydroxy-3-[4-[2-(1-pyrolodinyl)ethoxy]thiophenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophenehydrochloride ##STR64##

mp 137°-140° C. (dec). ¹ H NMR (DMSO-d₆) δ 9.86 (s, 1H), 9.80 (s, 1H),7.46 (d, J=8.6 Hz, 2H), 7.42 (d, J=8.7 Hz, 1H), 7.29 (d, J=2.0 Hz, 1H),6.96 (d, J=8.7 Hz, 2H), 6.87-6.81 (m, 5H), 4.21 (bt, 2H), 3.53-3.41 (m,4H), 3.01 (m, 2H), 1.95-1.82 (m, 4H). FD mass spec: 464. Anal. Calcd.for C₂₆ H₂₅ NO₃ S₂.1.0HCl: C, 62.45; H, 5.24; N, 2.80. Found: C, 62.36;H, 5.37; N, 2.61.

EXAMPLE 406-Hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(phenyl)]benzo[b]thiophenehydrochloride ##STR65##

To a solution of6-hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophenehydrochloride (5.00 g, 10.0 mmol) in 100 mL of anhydrous methylenechloride at 0° C. under N₂ was added triethylamine (8.38 mL, 60.0 mmol)followed by triflouromethanesulfonic anhydride (1.69 mL, 10.0 mmol). Theresulting mixture was allowed to gradually warm to room temperature andstirred for 1.5 hours. The reaction was then quenched by pouring into200 mL of saturated sodium bicarbonate solution. The aqueous phase wasthen extracted with ethylacetate (3×100 mL). The organic layer was dried(sodium sulfate) and concentrated in vacuo to an oil. Chromatography(0-3% methanol/chloroform) provided 2.82 g (39%) of6-triflouromethanesulfonate-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-triflouromethanesulfonatephenyl)]benzo[b]thiophene,1.82 g (31%) of a 1:1 mixture of 6-triflouromethanesulfonate-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-phenyl)]benzo[b]thiophene and3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-triflouromethanesulfonatephenyl)]benzo[b]thiophene,and 1.48 g (36%) of recovered staring material as the free base.

To a solution of a 1:1 mixture of monotriflate derivatives from the lastreaction (0.50 g, 0.84 mmol) in 60 mL of ethanol-ethyl acetate (5:1) wasadded triethylamine (2.0 mL) and 5% palladium-on-carbon (0.50 g). Theresulting mixture was hydrogenated at 40 psi for 2 hours. The mixturewas then filtered through Celite® to remove the catalyst. The filtratewas concentrated to an oil. The resulting mixture of monohydroxyderivatives was dissolved in ethyl acetate from which3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)]benzo[b]thiophene precipitated. Thefiltrate consisted of a 4:1 mixture of monohydroxy derivatives where6-hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(phenyl)]benzo[b]thiophenewas the major component. The filtrate was concentrated in vacuo, and theresulting solid dissolved in minimal ethyl acetate and treated withethyl ether.hydrochloric acid. The resulting solid was recrystallizedfrom ethanol to give 69 mg (18%) of isomerically pure6-hydroxy-3-[4-[2-(1-piperidinyl)ethoxy]phenoxy]-2-(phenyl)]benzo[b]thiophenehydrochloride. mp 217°-219° C. ¹ H NMR (DMSO-d₆) δ 9.87 (s, 1H), 7.64(d, J=7.5 Hz, 2H), 7.39-7.26 (m, 4H), 7.10 (d, J=8.6 Hz, 1H), 6.89 (s,4H), 6.78 (dd, J=8.6, 2.0 Hz, 1H), 4.22 (bt, 2H), 3.39-3.37 (m, 4H),2.97-2.90 (m, 2H), 1.74-1.60 (m, 5H), 1.39 (m, 1H). FD mass spec: 446.Anal. Calcd. for C₂₇ H₂₇ NO₃ S.1.0HCl: C, 67.28; H, 5.86; N, 2.91.Found: C, 67.00; H, 5.59; N, 2.87.

TEST PROCEDURE General Preparation Procedure

In the examples illustrating the methods, a post-menopausal model wasused in which effects of different treatments upon circulating lipidswere determined.

Seventy-five day old female Sprague Dawley rats (weight range of 200 to225 g) were obtained from Charles River Laboratories (Portage, Mich.).The animals were either bilaterally ovariectomized (OVX) or exposed to aSham surgical procedure at Charles River Laboratories, and then shippedafter one week. Upon arrival, they were housed in metal hanging cages ingroups of 3 or 4 per cage and had ad libitum access to food (calciumcontent approximately 0.5%) and water for one week. Room temperature wasmaintained at 22.2° ±1.7° C. with a minimum relative humidity of 40%.The photoperiod in the room was 12 hours light and 12 hours dark.

Dosing Regimen Tissue Collection

After a one week acclimation period (therefore, two weeks post-OVX)daily dosing with test compound was initiated. 17α-ethynyl estradiol orthe test compound were given orally, unless otherwise stated, as asuspension in 1% carboxymethylcellulose or dissolved in 20%cyclodextrin. Animals were dosed daily for 4 days. Following the dosingregimen, animals were weighed and anesthetized with a ketamine: Xylazine(2:1, V:V) mixture and a blood sample was collected by cardiac puncture.The animals were then sacrificed by asphyxiation with CO₂, the uteruswas removed through a midline incision, and a wet uterine weight wasdetermined.

Cholesterol Analysis

Blood samples were allowed to clot at room temperature for 2 hours, andserum was obtained following centrifugation for 10 minutes at 3000 rpm.Serum cholesterol was determined using a Boehringer Mannhelm Diagnosticshigh performance cholesterol assay. Briefly the cholesterol was oxidizedto cholest-4-en-3-one and hydrogen peroxide. The hydrogen peroxide wasthen reacted with phenol and 4-aminophenazone in the presence ofperoxidase to produce a p-quinone imine dye, which was readspectrophotemetrically at 500 nm. Cholesterol concentration was thencalculated against a standard curve. The entire assay was automatedusing a Biomek Automated Workstation.

Uterine Eosinophil Peroxidase (EPO) Assay

Uteri were kept at 4° C. until time of enzymatic analysis. The uteriwere then homogenized in 50 volumes of 50 mM Tris buffer (pH-8.0)containing 0.005% Triton X-100. Upon addition of 0.01% hydrogen peroxideand 10 mM O-phenylenediamine (final concentrations) in Tris buffer,increase in absorbance was monitored for one minute at 450 nm. Thepresence of eosonophils in the uterus is an indication of estrogenicactivity of a compound. The maximal velocity of a 15 second interval wasdetermined over the initial, linear portion of the reaction curve.

Source of Compound:

17α-ethynyl estradiol was obtained from Sigma Chemical Co., St. Louis,Mo.

Influence of Formula I Compounds on Serum Cholesterol and Determinationof Agonist/Non-Agonist Activity

Data presented in Table 1 below show comparative results amongovariectomized rats, rats treated with 17α-ethynyl estradiol (EE₂ ; anorally available form of estrogen), and rats treated with certaincompounds of the present invention. Although EE₂ caused a decrease inserum cholesterol when orally administered at 0.1 mg/kg/day, it alsoexerted a stimulatory action on the uterus so that EE₂ uterine weightwas substantially greater than the uterine weight of ovariectomized testanimals. This uterine response to estrogen is well recognized in theart.

Not only did the compounds of the present invention generally reduceserum cholesterol compared to the ovariectomized control animals, bututerine weight was only minimally increased to slightly decreased withthe majority of the formula compounds tested. Compared to estrogeniccompounds known in the art, the benefit of serum cholesterol reductionwithout adversely affecting uterine weight is quite rare and desirable.

As is expressed in the below data, estrogenicity also was assessed byevaluating the adverse response of eosinophil infiltration into theuterus. The compounds of the present invention did not cause anyincrease in the number of eosinophils observed in the stromal layer ofovariectomized rats, while estradiol cause a substantial, expectedincrease in eosinophil infiltration.

The data presented in the Tables 1 below reflects the response of 5 to 6rats per treatment.

                  TABLE 1                                                         ______________________________________                                                          Uterine           Serum                                                       Weight    Uterine Cholesterol                                       Dose      (% increase                                                                             EPO     (% decrease                               Compound                                                                              mg/kg     vs. OVX)  (V. max)                                                                              vs. OVX)                                  ______________________________________                                        EE.sub.2                                                                              0.1       229.2     308.1   94.8                                      Example 3                                                                             0.01      29.1      1.8     50.6                                              0.1       55.4      4.8     47.8                                              1.0       61.9      5.4     49.2                                      Example 4                                                                             0.1       33.2      3.9     53.7                                              1.0       35.6      4.8     62.1                                              10.0      34.7      3.0     65.3                                      Example 5                                                                             0.1       66.7      7.2     67.2                                              1.0       106.9     54.6    67.7                                              10.0      109.8     59.4    60.2                                      Example 7                                                                             0.1       32.0      4.8     56.2                                              1.0       44.3      4.5     42.6                                              5.0       41.6      4.8     29.5                                      Example 10                                                                            0.1       19.7      12.0    50.2                                              1.0       18.4      17.7    59.0                                              10.0      13.3      4.8     38.9                                      Example 19                                                                            0.01      11.4      2.1     25.1                                              0.1       24.9      2.4     45.3                                              1.0       24.7      3.6     53.6                                      Example 20                                                                            0.01      16.9      0.9     29.4                                              0.05      40.9      3.0     35.9                                              0.1       30.6      3.0     58.7                                      Example 21                                                                            0.01      21.0      1.2     26.8                                              0.1       24.8      4.8     47.5                                              1.0       51.4      9.3     54.4                                      Example 23                                                                            0.01      21.6      3.3     36.2                                              0.1       33.4      84.3    47.2                                              1.0       148.9     150.6   66.1                                      Example 24                                                                            0.01      9.2       3.6     23.7                                              0.1       18.2      0.9     46.4                                              1.0       81.0      29.4    79.3                                      Example 25                                                                            0.01      5.4       3.0     13.1                                              0.1       16.7      3.3     67.6                                              1.0       96.6      36.0    73.9                                      Example 26                                                                            0.01      14.0      4.8     29.0                                              0.1       81.0      29.1    45.2                                              1.0       117.1     175.1   62.7                                      Example 27                                                                            0.01      2.2       3.3     12.2                                              0.1       49.2      4.8     50.8                                              1.0       86.4      52.5    76.5                                      Example 29                                                                            0.01      0.0       3.3     9.2                                               0.1       17.2      4.8     43.8                                              1.0       31.0      6.0     39.4                                      Example 30                                                                            0.01      43.8      3.6     12.6                                              0.1       80.5      88.5    43.8                                              1.0       74.8      94.5    67.4                                      Example 36                                                                            0.1       40.6      0.9     62.7                                              1.0       24.1      1.3     57.5                                              10.0      32.0      4.8     58.7                                      ______________________________________                                    

In addition to the demonstrated benefits of the compounds of the presentinvention, especially when compared to estradiol, the above data clearlydemonstrate that compounds of Formula I are not estrogen mimetics.Furthermore, no deleterious toxicological effects (survival) wereobserved with any treatment.

Osteoporosis Test Procedure

Following the General Preparation Procedure, infra, the rats weretreated daily for 35 days (6 rats per treatment group) and sacrificed bycarbon dioxide asphyxiation on the 36th day. The 35 day time period wassufficient to allow maximal reduction in bone density, measured asdescribed herein. At the time of sacrifice, the uteri were removed,dissected free of extraneous tissue, and the fluid contents wereexpelled before determination of wet weight in order to confirm estrogendeficiency associated with complete ovariectomy. Uterine weight wasroutinely reduced about 75% in response to ovariectomy. The uteri werethen placed in 10% neutral buffered formalin to allow for subsequenthistological analysis.

The right femurs were excised and digitilized x-rays generated andanalyzed by an image analysis program (NIH image) at the distalmetaphysis. The proximal aspect of the tibiae from these animals werealso scanned by quantitative computed tomography.

In accordance with the above procedures, compounds of the presentinvention and ethynyl estradiol (EE₂) in 20% hydroxypropylβ-cyclodextrin were orally administered to test animals. Distal femurmetaphysis data presented in Tables 2 and 3 below are the results offormula I compound treatments compared to intact and ovariectomized testanimals. Results are reported as the mean ±the standard error of themean.

                  TABLE 2                                                         ______________________________________                                                                Distal Femur Metaphysis                                                       (X-ray Image Analysis-                                Compound/Treatment                                                                           Dose/kg  Gray Score                                            ______________________________________                                        Sham (20% cyclodextrin)                                                                      --       27.2 ± 6.0                                         Overiectomy control                                                                          --       8.1 ± 1.8                                          (20% cyclodextrin)                                                            EE2            0.1 mg   11.5 ± 2.9*                                        Example 19     0.1 mg   14.7 ± 1.9                                                        1.0 mg   15.0 ± 3.5*                                                       10.0 mg  15.3 ± 4.0*                                        ______________________________________                                         *P <= 0.5 two tailed Student's T Test on raw data.                       

                  TABLE 3                                                         ______________________________________                                                                  Distal Femur                                                                  Metaphysis (X-ray                                                             Image Analysis-Gray                                 Compound/Treatment                                                                           Dose/kg    Score                                               ______________________________________                                        Sham (20% cyclodextrin)                                                                      --         31.1 ± 6.3                                       Overiectomy control                                                                          --          6.2 ± 1.4                                       (20% cyclodextrin)                                                            EE2            0.1 mg     17.8 ± 3.5                                       Example 10     0.1 mg     15.3 ± 3.0                                                      1.0 mg     15.2 ± 3.7                                                      3.0 mg      18.5 ± 3.2*                                     Example 24     0.1 mg      18.3 ± 2.6*                                                    1.0 mg      19.6 ± 2.3*                                                    3.0 mg     17.1 ± 5.5                                       ______________________________________                                         *P <= 0.05 two tailed Student's T Test on raw data.                      

In summary, ovariectomy of the test animals caused a significantreduction in femur density compared to intact, vehicle treated controls.Orally administered ethynyl estradiol (EE₂) prevented this loss, but therisk of uterine stimulation with this treatment is ever-present.

The compounds of the present invention also prevented bone loss in ageneral, dose-dependent manner. Accordingly, the compounds of thepresent invention are useful for the treatment of post-menopausalsyndrome, particularly osteoporosis.

MCF-7 Proliferation Assay

MCF-7 breast adenocarcinoma cells (ATCC HTB 22) were maintained in MEM(minimal essential medium, phenol red-free, Sigma, St. Louis, Mo.)supplimented with 10% fetal bovine serum (FBS) (V/V), L-glutamine (2mM), sodium pyruvate (1 mM), HEPES{(N-[2-hydroxyethyl]piperazine-N'-[2-ethanesulfonic acid]10 mM},non-essential amino acids and bovine insulin (1 ug/mL) (maintenancemedium). Ten days prior to assay, MCF-7 cells were switched tomaintenance medium supplemented with 10% dextran coated charcoalstripped fetal bovine serum (DCC-FBS) assay medium) in place of 10% FBSto deplete internal stores of steroids. MCF-7 cells were removed frommaintenance flasks using cell dissociation medium (Ca++/Mg++ free HBSS(phenol red-free) supplemented with 10 mM HEPES and 2 mM EDTA). Cellswere washed twice with assay medium and adjusted to 80,000 cells/mL.Approximately 100 μL (8,000 cells) were added to flat-bottommicroculture wells (Costar 3596) and incubated at 37° C. in a 5% CO.sub.2 humidified incubator for 48 hours to allow for cell adherence andequilibration after transfer. Serial dilutions of drugs or DMSO as adiluent control were prepared in assay medium and 50 μL transferred totriplicate microcultures followed by 50 μL assay medium for a finalvolume of 200 μL. After an additional 48 hours at 37° C. in a 5% CO₂humidified incubator, microcultures were pulsed with tritiated thymidine(1 uCi/well) for 4 hours. Cultures were terminated by freezing at -70°C. for 24 hours followed by thawing and harvesting of microculturesusing a Skatron Semiautomatic Cell Harvester. Samples were counted byliquid scintillation using a Wallac BetaPlace β counter. Results inTable 4 below show the IC₅₀ for certain compounds of the presentinvention.

                  TABLE 4                                                         ______________________________________                                               Compound                                                                              IC.sub.50 nM                                                   ______________________________________                                               Example 3                                                                             4.0                                                                   Example 10                                                                            2.00                                                                  Example 19                                                                             0.028                                                                Example 21                                                                            0.05                                                                  Example 23                                                                            0.08                                                                  Example 36                                                                            0.28                                                           ______________________________________                                    

DMBA-Induced Mammary Tumor Inhibition

Estrogen-dependent mammary tumors are produced in female Sprague-Dawleyrats which are purchased from Harlan Industries, Indianapolis, Ind. Atabout 55 days of age, the rats receive a single oral feeding of 20 mg of7,12-dimethylbenz[a]anthracene (DMBA). About 6 weeks after DMBAadministration, the mammary glands are palpated at weekly intervals forthe appearance of tumors. Whenever one or more tumors appear, thelongest and shortest diameters of each tumor are measured with a metriccaliper, the measurements are recorded, and that animal is selected forexperimentation. An attempt is made to uniformly distribute the varioussizes of tumors in the treated and control groups such thataverage-sized tumors are equivalently distributed between test groups.Control groups and test groups for each experiment contain 5 to 9animals.

Compounds of Formula I are administered either through intraperitonealinjections in 2% acacia, or orally. Orally administered compounds areeither dissolved or suspended in 0.2 mL corn oil. Each treatment,including acacia and corn oil control treatments, is administered oncedaily to each test animal. Following the initial tumor measurement andselection of test animals, tumors are measured each week by theabove-mentioned method. The treatment and measurements of animalscontinue for 3 to 5 weeks at which time the final areas of the tumorsare determined. For each compound and control treatment, the change inthe mean tumor area is determined.

Uterine Fibrosis Test Procedures

Test 1

Between 3 and 20 women having uterine fibrosis are administered acompound of the present invention. The amount of compound administeredis from 0.1 to 1000 mg/day, and the period of administration is 3months.

The women are observed during the period of administration, and up to 3months after discontinuance of administration, for effects on uterinefibrosis.

Test 2

The same procedure is used as in Test 1, except the period ofadministration is 6 months.

Test 3

The same procedure is used as in Test 1, except the period ofadministration is 1 year.

Test 4

A. Induction of Fibroid Tumors in Guinea Pig

Prolonged estrogen stimulation is used to induce leiomyomata in sexuallymature female guinea pigs. Animals are dosed with estradiol 3-5 timesper week by injection for 2-4 months or until tumors arise. Treatmentsconsisting of a compound of the invention or vehicle is administereddaily for 3-16 weeks and then animals are sacrificed and the uteriharvested and analyzed for tumor regression.

B. Implantation of Human Uterine Fibroid Tissue in Nude Mice

Tissue from human leiomyomas are implanted into the peritoneal cavityand or uterine myometrium of sexually mature, castrated, female, nudemice. Exogenous estrogen are supplied to induce growth of the explantedtissue. In some cases, the harvested tumor cells are cultured in vitroprior to implantation. Treatment consisting of a compound of the presentinvention or vehicle is supplied by gastric lavage on a daily basis for3-16 weeks and implants are removed and measured for growth orregression. At the time of sacrifice, the uteri is harvested to assessthe status of the organ.

Test 5

A. Tissue from Human Uterine Fibroid Tumors is Harvested and Maintained,In Vitro, as Primary Nontransformed Cultures

Surgical specimens are pushed through a sterile mesh or sieve, oralternately teased apart from surrounding tissue to produce a singlecell suspension. Cells are maintained in media containing 10% serum andantibiotic. Rates of growth in the presence and absence of estrogen aredetermined. Cells are assayed for their ability to produce complementcomponent C3 and their response to growth factors and growth hormone. Invitro cultures are assessed for their proliferative response followingtreatment with progestins, GnRH, a compound of the present invention andvehicle. Levels of steroid hormone receptors are assessed weekly todetermine whether important cell characteristics are maintained invitro. Tissue from 5-25 patients are utilized.

Activity in at least one of the above tests indicates the compounds ofthe present invention are of potential in the treatment of uterinefibrosis.

Endometriosis Test Procedure

In Tests 1 and 2, effects of 14-day and 21-day administration ofcompounds of the present invention on the growth of explantedendometrial tissue can be examined.

Test 1

Twelve to thirty adult CD strain female rats are used as test animals.They are divided into three groups of equal numbers. The estrous cycleof all animals is monitored. On the day of proestrus, surgery isperformed on each female. Females in each group have the left uterinehorn removed, sectioned into small squares, and the squares are looselysutured at various sites adjacent to the mesenteric blood flow. Inaddition, females in Group 2 have the ovaries removed.

On the day following surgery, animals in Groups 1 and 2 receiveintraperitoneal injections of water for 14 days whereas animals in Group3 receive intraperitoneal injections of 1.0 mg of a compound of thepresent invention per kilogram of body weight for the same duration.Following 14 days of treatment, each female is sacrificed and theendometrial explants, adrenals, remaining uterus, and ovaries, whereapplicable, are removed and prepared for histological examination. Theovaries and adrenals are weighed.

Test 2

Twelve to thirty adult CD strain female rats are used as test animals.They are divided into two equal groups. The estrous cycle of all animalsis monitored. On the day of proestrus, surgery is performed on eachfemale. Females in each group have the left uterine horn removed,sectioned into small squares, and the squares are loosely sutured atvarious sites adjacent to the mesenteric blood flow.

Approximately 50 days following surgery, animals assigned to Group 1receive intraperitoneal injections of water for 21 days whereas animalsin Group 2 receive intraperitoneal injections of 1.0 mg of a compound ofthe present invention per kilogram of body weight for the same duration.Following 21 days of treatment, each female is sacrificed and theendometrial explants and adrenals are removed and weighed. The explantsare measured as an indication of growth. Estrous cycles are monitored.

Test 3

A. Surgical Induction of Endometriosis

Autographs of endometrial tissue are used to induce endometriosis inrats and/or rabbits. Female animals at reproductive maturity undergobilateral oophorectomy, and estrogen is supplied exogenously thusproviding a specific and constant level of hormone. Autologousendometrial tissue is implanted in the peritoneum of 5-150 animals andestrogen supplied to induce growth of the explanted tissue. Treatmentconsisting of a compound of the present invention is supplied by gastriclavage on a daily basis for 3-16 weeks, and implants are removed andmeasured for growth or regression. At the time of sacrifice, the intacthorn of the uterus is harvested to assess status of endometrium.

B. Implantation of Human Endometrial Tissue in Nude Mice

Tissue from human endometrial lesions is implanted into the peritoneumof sexually mature, castrated, female, nude mice. Exogenous estrogen issupplied to induce growth of the explanted tissue. In some cases, theharvested endometrial cells are cultured in vitro prior to implantation.Treatment consisting of a compound of the present invention supplied bygastric lavage on a daily basis for 3-16 weeks, and implants are removedand measured for growth or regression. At the time of sacrifice, theuteri is harvested to assess the status of the intact endometrium.

Test 4

A. Tissue from Human Endometrial Lesions is Harvested and Maintained InVitro as Primary Nontransformed Cultures

Surgical specimens are pushed through a sterile mesh or sieve, oralternately teased apart from surrounding tissue to produce a singlecell suspension. Cells are maintained in media containing 10% serum andantibiotic. Rates of growth in the presence and absence of estrogen aredetermined. Cells are assayed for their ability to produce complementcomponent C3 and their response to growth factors and growth hormone. Invitro cultures are assessed for their proliferative response followingtreatment with progestins, GnRH, a compound of the invention, andvehicle. Levels of steroid hormone receptors are assessed weekly todetermine whether important cell characteristics are maintained invitro. Tissue from 5-25 patients is utilized.

Activity in any of the above assays indicates that the compounds of thepresent invention are useful in the treatment of endometriosis.

Inhibition of Aortal Smooth Cell Proliferation/Restenosis Test Procedure

Compounds of the present invention have capacity to inhibit aortalsmooth muscle cell proliferation. This can be demonstrated by usingcultured smooth cells derived from rabbit aorta, proliferation beingdetermined by the measurement of DNA synthesis. Cells are obtained byexplant method as described in Ross, J. of Cell Bio. 50:172 (1971).Cells are plated in 96 well microtiter plates for five days. Thecultures become confluent and growth arrested. The cells are thentransferred to Dulbecco's Modified Eagle's Medium (DMEM) containing0.5-2% platelet poor plasma, 2 mM L-glutamine, 100 U/ml penicillin, 100mg ml streptomycin, 1 mC/ml ³ H-thymidine, 20 ng/ml platelet-derivedgrowth factor, and varying concentrations of the present compounds.Stock solution of the compounds is prepared in dimethyl sulphoxide andthen diluted to appropriate concentration (0.01-30 mM) in the aboveassay medium. Cells are then incubated at 37° C. for 24 hours under 5%CO₂ /95% air. At the end of 24 hours, the cells are fixed in methanol. ³H thymidine incorporation in DNA is then determined by scintillationcounting as described in Bonin, et al., Exp. Cell Res. 181: 475-482(1989).

Inhibition of aortal smooth muscle cell proliferation by the compoundsof the present invention are further demonstrated by determining theireffects on exponentially growing cells. Smooth muscle cells from rabbitaortae are seeded in 12 well tissue culture plates in DMEM containing10% fetal bovine serum, 2 mM L-glutamine, 100 U/ml penicillin, and 100mg/ml streptomycin. After 24 hours, the cells are attached and themedium is replaced with DMEM containing 10% serum, 2 mM L-glutamine, 100U/ml penicillin, 100 mg/ml streptomycin, and desired concentrations ofthe compounds. Cells are allowed to grow for four days. Cells aretreated with trypsin and the number of cells in each culture isdetermined by counting using a ZM-Coulter counter.

Activity in the above tests indicates that the compounds of the presentinvention are of potential in the treatment of restenosis.

The present invention also provides a method of alleviatingpost-menopausal syndrome in women which comprises the aforementionedmethod using compounds of Formula I and further comprises administeringto a woman an effective amount of estrogen or progestin. Thesetreatments are particularly useful for treating osteoporosis andlowering serum cholesterol because the patient will receive the benefitsof each pharmaceutical agent while the compounds of the presentinvention would inhibit undesirable side-effects of estrogen andprogestin. Activity of these combination treatments in any of thepost-menopausal tests, infra, indicates that the combination treatmentsare useful for alleviating the symptoms of post-menopausal symptoms inwomen.

Various forms of estrogen and progestin are commercially available.Estrogen-based agents include, for example, ethynyl estrogen (0.01-0.03mg/day), mestranol (0.05-0.15 mg/day), and conjugated estrogenichormones such as Premarin® (Wyeth-Ayerst; 0.3-2.5 mg/day).Progestin-based agents include, for example, medroxyprogesterone such asProvera® (Upjohn; 2.5-10 mg/day), norethylnodrel (1.0-10.0 mg/day), andnonethindrone (0.5-2.0 mg/day). A preferred estrogen-based compound isPremarin, and norethylnodrel and norethindrone are preferredprogestin-based agents.

The method of administration of each estrogen- and progestin-based agentis consistent with that which is known in the art. For the majority ofthe methods of the present invention, compounds of Formula I areadministered continuously, from 1 to 3 times daily. However, cyclicaltherapy may especially be useful in the treatment of endometriosis ormay be used acutely during painful attacks of the disease. In the caseof restenosis, therapy may be limited to short (1-6 months) intervalsfollowing medical procedures such as angioplasty.

As used herein, the term "effective amount" means an amount of compoundof the present invention which is capable of alleviating the symptoms ofthe various pathological conditions herein described. The specific doseof a compound administered according to this invention will, of course,be determined by the particular circumstances surrounding the caseincluding, for example, the compound administered, the route ofadministration, the state of being of the patient, and the pathologicalcondition being treated. A typical daily dose will contain a nontoxicdosage level of from about 5 mg to about 600 mg/day of a compound of thepresent invention. Preferred daily doses generally will be from about 15mg to about 80 mg/day.

The compounds of this invention can be administered by a variety ofroutes including oral, rectal, transdermal, subucutaneus, intravenous,intramuscular, and intranasal. These compounds preferably are formulatedprior to administration, the selection of which will be decided by theattending physician. Thus, another aspect of the present invention is apharmaceutical composition comprising an effective amount of a compoundof Formula I, or a pharmaceutically acceptable salt thereof, optionallycontaining an effective amount of estrogen or progestin, and apharmaceutically acceptable carrier, diluent, or excipient.

The total active ingredients in such formulations comprises from 0.1% to99.9% by weight of the formulation. By "pharmaceutically acceptable" itis meant the carrier, diluent, excipients and salt must be compatiblewith the other ingredients of the formulation, and not deleterious tothe recipient thereof.

Pharmaceutical formulations of the present invention can be prepared byprocedures known in the art using well known and readily availableingredients. For example, the compounds of formula I, with or without anestrogen or progestin compound, can be formulated with commonexcipients, diluents, or carriers, and formed into tablets, capsules,suspensions, powders, and the like. Examples of excipients, diluents,and carriers that are suitable for such formulations include thefollowing: fillers and extenders such as starch, sugars, mannitol, andsilicic derivatives; binding agents such as carboxymethyl cellulose andother cellulose derivatives, alginates, gelatin, andpolyvinylpyrrolidone; moisturizing agents such as glycerol;disintegrating agents such as calcium carbonate and sodium bicarbonate;agents for retarding dissolution such as paraffin; resorptionaccelerators such as quaternary ammonium compounds; surface activeagents such as cetyl alcohol, glycerol monostearate; adsorptive carrierssuch as kaolin and bentonite; and lubricants such as talc, calcium andmagnesium stearate, and solid polyethyl glycols.

The compounds also can be formulated as elixirs or solutions forconvenient oral administration or as solutions appropriate forparenteral administration, for example, by intramuscular, subcutaneousor intravenous routes. Additionally, the compounds are well suited toformulation as sustained release dosage forms and the like. Theformulations can be so constituted that they release the activeingredient only or preferably in a particular physiological location,possibly over a period of time. The coatings, envelopes, and protectivematrices may be made, for example, from polymeric substances or waxes.

Compounds of formula I, alone or in combination with a pharmaceuticalagent of the present invention, generally will be administered in aconvenient formulation. The following formulation examples only areillustrative and are not intended to limit the scope of the presentinvention.

FORMULATIONS

In the formulations which follow, "active ingredient" means a compoundof formula I, or a salt or solvate thereof.

Formulation 1: Gelatin Capsules

    ______________________________________                                        Hard gelatin capsules are                                                     prepared using the                                                            Ingredient        Quantity (mg/capsule)                                       ______________________________________                                        Active ingredient 0.1-1000                                                    Starch, NF        0-650                                                       Starch flowable powder                                                                          0-650                                                       Silicone fluid 350 centistokes                                                                  0-15                                                        ______________________________________                                    

The formulation above may be changed in compliance with the reasonablevariations provided.

A tablet formulation is prepared using the ingredients below:

Formulation 2: Tablets

    ______________________________________                                        Ingredient       Quantity (mg/tablet)                                         ______________________________________                                        Active ingredient                                                                                2.5-1000                                                   Cellulose, microcrystalline                                                                    200-650                                                      Silicon dioxide, fumed                                                                          10-650                                                      Stearate acid     5-15                                                        ______________________________________                                    

The components are blended and compressed to form tablets.

Alternatively, tablets each containing 2.5-1000 mg of active ingredientare made up as follows:

Formulation 3: Tablets

    ______________________________________                                        Ingredient          Quantity (mg/tablet)                                      ______________________________________                                        Active ingredient   25-1000                                                   Starch              45                                                        Cellulose, microcrystalline                                                                       35                                                        Polyvinylpyrrolidone                                                                               4                                                        (as 10% solution in water)                                                    Sodium carboxymethyl cellulose                                                                     4.5                                                      Magnesium stearate   0.5                                                      Talc                 1                                                        ______________________________________                                    

The active ingredient, starch, and cellulose are passed through a No. 45mesh U.S. sieve and mixed thoroughly. The solution ofpolyvinylpyrrolidone is mixed with the resultant powders which are thenpassed through a No. 14 mesh U.S. sieve. The granules so produced aredried at 50°-60° C. and passed through a No. 18 mesh U.S. sieve. Thesodium carboxymethyl starch, magnesium stearate, and talc, previouslypassed through a No. 60 U.S. sieve, are then added to the granuleswhich, after mixing, are compressed on a tablet machine to yieldtablets.

Suspensions each containing 0.1-1000 mg of medicament per 5 ml dose aremade as follows:

Formulation 4: Suspensions

    ______________________________________                                        Ingredient           Quantity (mg/5 ml)                                       ______________________________________                                        Active ingredient    0.1-1000 mg                                              Sodium carboxymethyl cellulose                                                                     50 mg                                                    Syrup                1.25 mg                                                  Benzoic acid solution                                                                              0.10 mL                                                  Flavor               q.v.                                                     Color                q.v.                                                     Purified water to    5 mL                                                     ______________________________________                                    

The medicament is passed through a No. 45 mesh U.S. sieve and mixed withthe sodium carboxymethyl cellulose and syrup to form a smooth paste. Thebenzoic acid solution, flavor, and color are diluted with some of thewater and added, with stirring. Sufficient water is then added toproduce the required volume.

An aerosol solution is prepared containing the following ingredients:

Formulation 5: Aerosol

    ______________________________________                                        Ingredient           Quantity (% by weight)                                   ______________________________________                                        Active ingredient    0.25                                                     Ethanol              25.75                                                    Propellant 22 (Chlorodifluoromethane)                                                              70.00                                                    ______________________________________                                    

The active ingredient is mixed with ethanol and the mixture added to aportion of the propellant 22, cooled to 30° C., and transferred to afilling device. The required amount is then fed to a stainless steelcontainer and diluted with the remaining propellant. The valve units arethen fitted to the container.

Suppositories are prepared as follows:

Formulation 6: Suppositories

    ______________________________________                                        Ingredient         Quantity (mg/suppository)                                  ______________________________________                                        Active ingredient    250                                                      Saturated fatty acid qlycerides                                                                  2,000                                                      ______________________________________                                    

The active ingredient is passed through a No. 60 mesh U.S. sieve andsuspended in the saturated fatty acid glycerides previously melted usingthe minimal necessary heat. The mixture is then poured into asuppository mold of nominal 2 g capacity and allowed to cool.

An intravenous formulation is prepared as follows:

Formulation 7: Intravenous Solution

    ______________________________________                                        Ingredient            Quantity                                                ______________________________________                                        Active ingredient     50 mg                                                   Isotonic saline       1,000 mL                                                ______________________________________                                    

The solution of the above ingredients is intravenously administered to apatient at a rate of about 1 mL per minute.

Formulation 8: Combination Capsule I

    ______________________________________                                        Ingredient     Quantity (mg/capsule)                                          ______________________________________                                        Active ingredient                                                                            50                                                             Premarin       1                                                              Avicel pH 101  50                                                             Starch 1500    117.50                                                         Silicon Oil    2                                                              Tween 80       0.50                                                           Cab-O-Sil      0.25                                                           ______________________________________                                    

Formulation 9: Combination Capsule II

    ______________________________________                                        Ingredient     Quantity (mg/capsule)                                          ______________________________________                                        Active ingredient                                                                            50                                                             Norethylnodrel 5                                                              Avicel pH 101  82.50                                                          Starch 1500    90                                                             Silicon Oil    2                                                              Tween 80       0.50                                                           ______________________________________                                    

Formulation 10: Combination Tablet

    ______________________________________                                        Ingredient     Quantity (mg/capsule)                                          ______________________________________                                        Active ingredient                                                                            50                                                             Premarin       1                                                              Corn Starch NF 50                                                             Povidone, K29-32                                                                             6                                                              Avicel pH 101  41.50                                                          Avicel pH 102  136.50                                                         Crospovidone XL10                                                                            2.50                                                           Magnesium Stearate                                                                           0.50                                                           Cab-O-Sil      0.50                                                           ______________________________________                                    

I claim:
 1. A method for inhibiting restenosis comprising administeringto a human in need of treatment an effective amount of a compound offormula I ##STR66## wherein R¹ is --H, --OH, --O(C₁ -C₄ alkyl), --OCOC₆H₅, --OCO(C₁ -C₆ alkyl), or --OSO₂ (C₂ -C₆ alkyl);R² is --H, --OH,--O(C₁ -C₄ alkyl), --OCOC₆ H₅, --OCO(C₁ -C₆ alkyl), --OSO₂ (C₂ -C₆alkyl), or halo, providing when Z is --S--, R² is not halo; R³ is1-piperidinyl, 1-pyrrolidinyl, methyl-1-pyrrolidinyl,dimethyl-1-pyrrolidinyl, 4-morpholino, dimethylamino, diethylamino,diisopropylamino, or 1-hexamethyleneimino; n is 2 or 3; and z is --O--or --S--;or a pharmaceutically acceptable salt thereof.